Handbook of Sugar A 00 Browne

A HANDBOOK OF
SUGAR ANALYSIS
A PRACTICAL AND DESCRIPTIVE TREATISE FOR USE
IN
RESEARCH, TECHNICAL AND CONTROL LABORATORIES
BY
C.
A.
BROWNE,
New
PH.D.
York Sugar Trade Laboratory {Formerly Chief of the Sugar Laboratory, U. 8. Bureau of Chemistry, Washington, D.C., and Research Chemist of the Louisiana
Chemist in charge of the
Sugar Experiment Station, New Orleans,
La.)
SECOND EDITION
FIRST THOUSAND
NEW YORK JOHN WILEY & SONS

LONDON:
CHAPMAN & HALL,

1912
LIMITED
COPYRIGHT, 1912, BY
C. A.
BROWNE
Copyright, 1912, in Great Britain
Stanbepe iprcss
F.
H.
GILSON COMPANY
BOSTON,
U.S.A.
TP.3Z2
AGRIC.
LIBRARY
DEDICATED
TO HIS TEACHER,
GEH.-RATH PROF. DR.

AS A TOKEN OF GRATITUDE
B.
TOLLENS,
OF GOTTINGEN UNIVERSITY,
AND ESTEEM,
BY THE AUTHOE
PREFACE
THE subject of sugar analysis, which a generation ago was limited to determinations of density, specific rotation and reducing power,
ments
has greatly expanded within the past twenty-five years. Instruof greater accuracy have been devised, old methods have been improved and new methods have been discovered. In the present
volume the purpose of the author has been to give a rather wide, but a by no means complete, selection of the more recent methods of sugar analysis and at the same time to retain the more important
features of the older textbooks.
The range
of sugar analysis
is
so broad that in the selection of
methods the author has been guided largely by his own experience in various research, technical and control laboratories. While the particular methods chosen for description may not in all cases meet with
it is hoped that the underlying principles of sugar have been covered sufficiently to enable the chemist to make analysis his own applications and modifications. References to special works and original articles will assist the chemist in case he desires to follow
general approval
some special line of investigation more fully. Next to the knowledge of a method the most important fact which the student of sugar analysis must acquire is the knowledge of this
method's limitations. The great susceptibility of the sugars to chemical changes and to variations in specific rotation, reducing power and other "constants" is a factor which the sugar chemist must always bear in mind. The prescribed methods of analysis are usually
too silent upon these points, and the inexperienced chemist often proceeds to make general use of a formula or method which has only a limited applicability. The author has endeavored to correct this tendency by including with the description of each method a brief
account of its applicability and limitations. In the examination of sugar-containing materials the problems of analysis are much simplified by a knowledge of what one may expect to find. The author has felt that a work upon sugar analysis is not In of the sugars themselves. complete without some
description
Part II of the present volume, he has therefore included a brief
Vi
PREFACE
account of the occurrence, methods of preparation, properties and

Brief reactions of the different sugars and their allied derivatives. references are also made to methods of sugar synthesis; the latter play such an important part in the separation and isolation of the rarer
is not fully equipped without some knowledge of synthetic processes. The principal textbooks and journals which have been consulted in preparing the present volume are named in the Bibliography. The author's obligations to these are indicated in most cases by the
sugars that the sugar analyst
In reviewing original papers, the abstracts and references contained in Lippmann's "Chemie der Zuckerarten" and his "Berichte liber die wichtigsten Arbeiten aus dem Gebiete der reinen Zuckerfootnotes.
chemie," published semiannually in "Die Deutsche Zuckerindustrie," have been of invaluable service. In concluding his task, which has extended with many interruptions over a period of five years, the author desires to thank the many friends and coworkers who, by their help and encouragement, have
greatly lightened his labors. Special obligations are due to Dr. C. S.
section
Hudson for reviewing the and to Prof. H. C. mutarotation Sherman for suggestions upon methods for diastatic determining upon power. Acknowledgement is also made of courtesies extended by Mr. A. H. Bryan and by Mr.
G.
W.
Rolfe.
For the use of cuts contained in Dr. G. L. Spencer's "Handbook for Cane Sugar Manufacturers" and in A. E. Leach's "Food Inspection and Analysis" the author owes an acknowledgment to the authors of these books and to his publishers Messrs. John Wiley & Sons. To the latter also he would express his appreciation of the hearty support which has been given and of the generous consideration which has been shown for the many delays incident to the completion of the work.
NEW YORK,
N. Y., August, 1912.
BIBLIOGRAPHY
SPECIAL
Author or Editor
WORKS
Title
Abderhalden Abderhalden
Allen
Biochemisches Handlexikon, Vol. II (1911). Handbuch der Biochemischen Arbeitsmethoden, Vol II (1909).. Commercial Organic Analysis, Vol. I
(1901).
Armstrong
Basset
The Simple Carbohydrates and

sides (1910).
the Gluco-
Guide pratique du Fabricant

(1872).
. .
de
Sucre
Browne
Bryan ....
Chemical Analysis and Composition of American Honeys (1908). Bull. 110, U. S. Bureau of Chemistry. Analyses of Sugar Beets, 1905 to 1910, together with Methods of Sugar Determination (1911). Bull. 146, U. S. Bureau of Chemistry. Hilfsbuch fur Nahrungsmittelchemiker
(1900).
Bujard and Baier

Claassen (Hall and Rolfe) Cross and Bevan Cross and Bevan
Beet Sugar Manufacture (1906).

Cellulose (1895). Researches on Cellulose, 1895-1900 (1901). Biochemie der Pflanzen, Vol. I (1905). Cane Sugar (1911). Die Zersetzung stickstofffreier organischer Substanzen durch Bakterien (1902).
Czapek
Deerr
Emmerling
Fischer,
Fischer, F.
Untersuchungen uber Kohlenhydrate und Fermente (1884-1908), (1909). Handbuch der chemischen Technologic,
Vol. II (1902). L' Analyse chimique en Sucreries et Raffineries de Cannes et Betteraves (1907). Anleitung zur Untersuchung der fiir die Zuckerindustrie in Betracht kommenden Rohmaterialien, Produkte, Nebenprodukte und Hilfssubstanzen (1903). Lehrbuch der Angewandten Optik in der Chemie. Spectralanalyse, Mikroskopie,
Fribourg
Friihling....
Gange
Polarisation (1886).
Geerligs Geerligs
. .
Cane Sugar and its Manufacture Methods of Chemical Control
(1909). in Cane
Gredinger Jago Konig....

Lafar.
.
.
Sugar Factories (1905). Die Raffination des Zuckers (1909).
The Technology
of Bread Making (1911). Die Untersuchung landwirtschaftlich und
Landolt
gewerblich wichtiger Stoffe (1898). Technische Mykologie (1897-1907). Das optische Drehungsvermogen organischer Substanzen und dessen praktische Anwendungen (1898). Food Inspection and Analysis (1911). Die Chemie der Zuckerarten (1904).
Leach
Lippmann
i^
Vlll
BIBLIOGRAPHY
Author or Editor
Title
Maquenne
Mittelstaedt (Bourbakis)
Les
Sucres
(1900).
et
leurs
principaux
Derives
Technical Calculations for Sugar Works

(1910).
Pavy
Plimmer
Preston Rolfe..
Riimpler.
Sidersky Sidersky
,
Sherman ....
Physiology of the Carbohydrates (1894). The Chemical Changes and Products Resulting from Fermentations (1903). The Theory of Light (1901). The Polariscope in the Chemical Laboratory (1905). Die Nichtzuckerstoffe der Ruben (1898). Methods of Organic Analysis (1912). Les Densites des Solutions Sucrees a differentes Temperatures (1908). Sucrerie, de Raffinerie et de Glucoserie (1909). Polarisation et Saccharimetrie (1908). Handbook for Cane-Sugar Manufacturers and their Chemists (1906). The Principles and Practice of Brewing
Manuel du Chimiste de
Sidersky,
Spencer
Sykes and Ling.
(1907).
Tervooren
Methoden van Onderzoek der bij de Java Rietsuiker-Industrie voorkomende Producten (1908). Kurzes Handbuch
der
Tollens.
Kohlenhydrate
Tucker
Van't Hoff (Marsh)
.
.
.
Walker Ware..
of Sugar Analysis (1905). Chemistry in Space (1891) Introduction to Physical Chemistry (1903). Beet Sugar Manufacture and Refining
A Manual
(1895-8).
(1905-7).
Wein (Frew)
Wiechmann Wiedemann and Ebert

Wiley....
Wiley..
Tables for the Quantitative Estimation of the Sugars (1896). Sugar Analysis (1898). Physikalisches Praktikum mit besonderer der Beriicksichtigung Physikalisch-
The
Chemischen Methoden (1899). Principles and Practice of Agricultural
Official
Analysis, Vol. Ill (1897). and Provisional Methods of Analysis, Association of Official Agricultural
Chemists.
Bureau
Bull. 107 of Chemistry.
(Revised) U. S.
PERIODICALS
Abbreviation
Title
Am. Chem. Jour Am. Sugar Ind

Analyst
American Chemical Journal. American Sugar Industry.

Analyst.
Ann
Annalen der Chemie
Ann. chim. phys Archief Java Suiker Ind.. Archiv Pharm.

.
.
(Liebig's).
Annales de chimie et de physique. Archief voor de Java Suiker Industrie. Archiv der Pharmazie. Berichte der deutschen chemischen Gesellschaft.
Biochem. Zeitschrift Bull, assoc. chim. sucr. dist.
Biochemische Zeitschrift. Bulletin de I'association des chimistes de sucrerie et de distillerie de France et

des colonies.
BIBLIOGRAPHY
Abbreviation
Bull. soc.
Title
chim
Centralblatt Centrbl. Zuckerind
Bulletin de la societe chimique de France. Chemisches Centralblatt. Centralblatt fur die Zuckerindustrie.
Chem. News
Chemiker-Ztg.
Chemical News and Journal of Physical

Science.
Compt. rend..
Deut. Zuckerind Dingier 's Poly tech. Jour. Int. Sugar Jour J. Am. Chem. Soc J. Chem. Soc J. fabr. sucre
Jour, f Landwirtsch J. Ind. Eng. Chem
.
Chemiker-Zeitung Comptes rendus hebdomadaires des seances de 1'academie des sciences. Die Deutsche Zuckerindustrie
.
J. J.
pharm
pharm. chim.
.
prakt. Chem. J. Soc. Chem. Ind.

J.
.
Dingier 's Polytechniches Journal. International Sugar Journal. Journal of the American Chemical Society. Journal of the Chemical Society (London). Journal des fabricants de sucre. Journal fur Landwirtschaft. Journal of Industrial and Engineering Chemistry. Journal de pharmacie. Journal de pharmacie et de chimie. Journal fur prakt ische Chemie. Journal of the Society of Chemical In-
The
dustry.
La. Planter
The Louisiana
ufacturer.
Planter-
and
Sugar ManVersuchs-Sta-
Land. Vers.-Stat..
Die
landwirthschaftlichen
tionen.
Monatshefte
Mon. Neue
scient Zeitschrif t Oest.-Ung. Z. Zuckerind.

Pfliiger's
Monatshefte fiir Chemie. Moniteur scientifique.
Archiv
Neue Zeitschrif t fiir Riibenzuckerindustrie. Oesterreichisch-Ungarische Zeitschrift fiir Zuckerindustrie und Landwirthschaft. Pfliiger's Archiv fiir die gesammte Physiologic der Menschen und der Thiere.
Poggendorff's Annalen. Proceedings of the Association of Official Agricultural Chemists. Proceedings of the International Congress of Applied Chemistry. Recueil des travaux chimiques des PaysBas. Sitzungsberichte der kaiserlichen Akademie der Wissenschaften, Wien. Stammer's Jahresbericht iiber die Untersuchungen und Fortschritte auf dem Gesamtgebiete der Zuckerfabrikation. La Sucrerie Beige. West Indian Bulletin. Wochenschrift fiir Brauerei.
Zeitschrift fiir analytische Chemie. Zeitschrift fiir angewandte Chemie. Zeitschrift fiir Instrumentenkunde. Zeitschrift fiir physikalische Chemie. Zeitschrift fiir physiologische Chemie. Zeitschrift fiir Spirit usindustrie. Zeitschrift fur Untersuchung der Nahrungs-
Pogg. Ann Proceedings A. O. A.
Proceedings Int. Cong. App. Chem.

Rec. trav. Pays-Bas
Sitzungsber. Wiener
Akad
. .
Stammer's Jahresbericht
Sucrerie Beige West Indian Bull Wochenschr. f. Brauerei.

Z. Z. Z. Z. Z. Z. Z.
analyt.
angew. Instrument
physik.
Chem Chem Chem
physiol. Chem Spiritusind
Unters. Nahr. Genussm.
und Genussmittel.
Z. Ver.
Deut. Zuckerind.
Zeitschrift
Z. Zuckerind.
Bohmen..
des Vereins der Deutschen Zuckerindustrie. Zeitschrift fiir Zuckerindustrie in Bohmen.
TABLE OF CONTENTS
PAGE
PREFACE
BIBLIOGRAPHY
v v jj
PART
CHAP.
I.
I
1
PHYSICAL AND CHEMICAL METHODS OF SUGAR ANALYSIS
SAMPLING OF SUGAR AND SUGAR PRODUCTS
II.
DETERMINATION OF MOISTURE IN SUGARS AND SUGAR PRODUCTS BY METHODS OF DRYING

DENSIMETRIC METHODS OF ANALYSIS
PRINCIPLE AND USES OF THE REFRACTOMETER
15
III.
27
50
.
IV.
V. POLARIZED LIGHT, VI.

VII.
VIII.
'
THEORY AND DESCRIPTION OF POLARIMETERS
76
THEORY AND DESCRIPTION OF SACCHARIMETERS

POLARISCOPE ACCESSORIES
SPECIFIC ROTATION OF SUGARS
108 146
172
194
IX.
METHODS OF SIMPLE POLARIZATION
X. METHODS OF INVERT OR DOUBLE POLARIZATION

XI. SPECIAL
XII. MISCELLANEOUS PHYSICAL TION OF SUGARS
XIII. QUALITATIVE
263
METHODS OF SACCHARIMETRY 287 METHODS AS APPLIED TO THE EXAMINA307 333

388
METHODS FOR THE IDENTIFICATION OF SUGARS XIV. REDUCTION METHODS FOR DETERMINING SUGARS XV. SPECIAL QUANTITATIVE METHODS XVI. COMBINED METHODS AND THE ANALYSIS OF SUGAR MIXTURES.
449
...
472
XVII. MISCELLANEOUS APPLICATIONS
494
PART
II
THE OCCURRENCE, METHODS OF PREPARATION, PROPERTIES AND PRINCIPAL

REACTIONS OF THE SUGARS AND ALLIED DERIVATIVES
XVIII.
CLASSIFICATION OF THE SUGARS AND THEIR FORMATION IN NATURE.
525
527 535
643
731
751
XIX. THE XX. THE XXI. THE XXII. THE
MONOSACCHARIDES
DISACCHARIDES
TRISACCHARIDES AND TETRASACCHARIDES
AMINO SUGARS AND THE CYCLOSES
XXIII. THE SUGAR ALCOHOLS AND SUGAR ACIDS
764
789
xiii
APPENDIX OF SUGAR TABLES

INDEX..
PAET
PHYSICAL AND CHEMICAL METHODS OF SUGAK ANALYSIS
ANALYSIS
CHAPTER
I

IN the analysis of sugars and sugar products, special stress must be upon the correctness of sample. Accuracy in analytical details is of no value unless the portion of substance weighed out for examination is an accurate sample of the entire lot of product in question. While the chemist is not always charged with the supervision of sampling, he should, nevertheless, acquaint himself so far as possible with the history of his product before it is received.. In this way he may often explain differences which might otherwise be attributed to mistakes of analysis. A few introductory pages devoted to the general subject of sampling may, therefore, not be amiss.
laid
The
nected therewith,
best illustration of methods of sampling, is furnished by raw cane sugar.

is
and
of the errors con-
commodity
selected first
and discussed
in
The sampling of this somewhat fuller detail.
SAMPLING OF
comes
RAW
SUGARS
countries
The raw sugar imported from the various sugar-producing
in a great variety of forms. Centrifugal sugar, from Cuba, Porto Rico, and most of the West Indian Islands, comes in 300-lb. jute bags; sugar from the Hawaiian Islands comes in 125-lb. bags;
sugar from Java comes either in bags or large cylindrical baskets weighing from 500 to 700 Ibs.; sugar from the Philippines comes in small wicker mats weighing about 50 Ibs.; Muscovado sugars, which
are purged
by draining and contain much
molasses,
come usually
in
large hogsheads.
In addition to the above forms of package, sugars

in boxes, barrels, grass mats, ceroons,
come occasionally
receptacles.
and other
The need for carefully prescribed rules in sampling sugar becomes at once self-evident when we consider the different forms of the package
and the exceedingly variable character
3
of the sugar
which
may
be con-
4
tained therein.
SUGAR ANALYSIS
The
sugar, for example,
may contain lumps of higher the product; the sugar may finer of the part or lower polarization than of molasses, sometimes as high as also retain considerable amounts " " foots or storage and form the transit 30 per cent, which drain during
bottom of the package. The difference in composition between the top and bottom layers of a hogshead of Muscovado sugar, which " foots" easily, is very marked. In addition to the is a kind that
at the
differences
the differences in composition

lot.
also
sugar within the single packages are between different packages of the same These differences may be the result of manufacture; they may result when no dunnage is used for covering the bottom of the
in composition of
Fig. 1.
Trier for sampling sugar.
holds of the ships used for transport, with the result that the bottom tiers of sugar may be damaged through absorption of bilge water. In
many
cases the top tiers of sugar suffer the damage, as when sugars sweat beneath the hatches; the vapors from the warm sugar rise, conIf the dense, and then drop back upon the upper layers of the cargo.
packages of sugar run unevenly
it is difficult
to secure a representative
The most approved method fraction unless every container is sampled. of sampling at present is to take a specimen of sugar so far as possible
from every package.* Sugar is sampled in the same
way
as fertilizers
and many other
commodities, by means of a trier. This implement (Fig. 1) consists of a long pointed rod of steel with a groove or spoon upon one side. A
* For a discussion of this and other points pertaining to methods of sampling raw sugar in different countries see paper by F. G. Wiechmann (Int. Sugar Journ., 9, 18-28) read before the Fifth Meeting of the International Commission for Uniform Methods of Sugar Analysis, Bern, 1906.

thrust of the trier into the package forces the sugar along its pathway tightly into the bowl of the spoon; the sugar thus adhering, after the trier is withdrawn, is removed by the thumb, or by means of a scraper,
and the process is continued until a sufficient number of packages have been sampled to constitute a mix; this number may vary, according to the size of lot and kind of sugar, from one package to several thousand. The practice of the New York Sugar Trade is to mix twice daily, and in no case is a sample to remain uninto a covered bucket,
mixed over night.

It is of course
important that the
triers of the different
workmen
are sampling a given lot of sugar should be exactly alike, esThe specifications pecially as regards the dimensions of the spoons.
of the
plicit
who
United States Treasury Department Regulations* are very exupon this point and give the following dimensions of the short,
barrel triers.
long,
and
TABLE
Giving Dimensions of Triers for Sampling Sugar
SUGAR ANALYSIS
kinds of sugar under certain atmospheric conditions of humidity. The surface of the metal should be smooth and bright; the United
States Treasury Regulations attach a penalty in case of samplers who When ready for making the composite sample, neglect this precaution.
bottles to receive the
after
tions.
the contents of the sugar bucket are thoroughly mixed; the cans and sample are compactly filled, labeled, and sealed,
which they are sent to the chemists who are to make the polariza-
The general rule in sampling sugar is that the package shall be stabbed at the middle to the center, and if this practice is conscientiously followed it will give no doubt as fair a sample as can be secured under the hurried conditions of discharging a cargo. There are times,
however, when it is impossible to follow this rule. Sugar which has remained for a long time in storage will sometimes solidify upon the approach of cold weather to a hard mass of material resembling concrete,
a circumstance due to the evaporation of moisture and cement-
trier is almost useless under these coning together of the grain. ditions and such sugar is rarely sampled properly. The sugar broken,
from the outside of the package is not a sometimes resorted to with hard sugar in order to open a passage for the trier; this is much better than just skimming the outside, but is far from satisfactory.
or chipped off, correct sample.
by the
trier
pickaxe
is
To eliminate so far as possible the errors of personal equation in sampling, the practice of the New York Sugar Trade is for the samplers of buyer and seller to work alternately hour by hour; the one party in
the interval of rest exercising a control upon the operations of the other. The tendencies to draw too high and too low from the package
are thus counterbalanced and the personal errors equalized. This method seems as good as any that can be devised. The liability of change in composition of the product during sampling is an exceedingly important factor in the valuation of any commodity, and more important perhaps in the case of sugar than almost any
other staple. Raw cane sugar upon exposure to the air may either absorb or lose moisture according to the conditions of atmospheric humidity. If the latter be very high or low, and the sugar be exposed to the air for any great length of time during drawing or mixing the
sample, a considerable error may be introduced into the composition of the product. The buckets, which hold the samples for mixing, should always be kept tightly covered; this precaution will reduce the
from absorption and evaporation to a large extent, although with present methods of sampling the errors from this source will never be
errors
completely eliminated. On rainy days sugar is rarely sampled at the pier, and this is a wise precaution, considering the rapidity with which sugar absorbs moisture from a saturated atmosphere. No matter how
pure the sugar, there will be absorption under such conditions, the amount of moisture taken up depending upon the initial dryness of the sugar, the fineness of the grain and the hygroscopic character of the im-
be placed in a dish over water under a closed soon absorb moisture enough to liquefy, and, according to the phase rule, this absorption of moisture will continue until the pressures of water vapor for solution and atmosphere are the same. Theoretically this limit is infinity, and if the dish under the bell jar be weighed from day to day it will be found that the liquefied sugar will continue to attract moisture as long as one cares to follow the experiment.
bell jar, it will
purities present. If a layer of sugar
If the atmosphere is not completely saturated, the absorption of moisture by the sugar is less rapid, and with further decrease in humidity a point of equilibrium is soon reached where there is neither absorption
This point of equilibrium, which represents equality vapor pressure between the moisture of the sugar and the air, is With still further decrease in humidity different for different sugars.
nor evaporation.
of
the sugar begins to give up moisture, the rate of loss increasing as the percentage of saturation in the air becomes less and less.
In the following table the percentages of moisture which different sugars gain or lose at 100 per cent relative humidity and at 60 per cent relative humidity are given, and the changes in moisture content at the point of equilibrium. Two grams of sugar were spread in a thin
layer upon a watch glass and the change in weight noted after regular intervals of time in one case over water under a bell jar, and in the
other case upon exposure to the open ments was 20 C.
air.
The temperature
of experi-
TABLE
II
Showing Variations in Moisture Content of Sugars
SUGAR ANALYSIS
After the point of equilibrium was reached upon exposure of the above sugars to the air, no change in weight was noted as long as the temperature and relative humidity remained unchanged; with fluctuations in the latter corresponding gains and losses were always observed
by sugars under excessive humidity, no relationship can be traced in the above table between composition and rate of absorption. The refined granulated sugar and the lowgrade mats have equally high absorptive powers and the high-grade Peruvian crystals and the Cuban molasses sugar equally low absorptive powers. If the grain of these sugars is compared, however, it will be seen that the Peruvian crystals and molasses sugar of low absorptive power have the largest grain and that the granulated sugar and mat sugar of highest absorptive power have the smallest grain, so that the
physical condition of the sugar is a very important factor in the influences which bear upon absorption. As to the evaporation of moisture from sugars under diminished
in the weight of the sugars. As to the absorption of moisture
humidity, the table shows a very definite relationship between compo-
and rate of evaporation, this rate being, as would be supposed, roughly proportional to the initial moisture content of the sugar. The percentage of residual moisture in a sugar at the point of equilibrium is a
sition
function of the hygroscopic power of the non-sugars, and with the sugars of lowest purity (highest molasses content).
sults
is
is
greatest
The point of greatest importance, in the bearing which these rehave upon the changes in composition of sugar during sampling,
moisture
that the gain or loss in weight through absorption or evaporation of is most rapid at the beginning. A comparison recently made
by the author of the changes in moisture content which sugars undergo upon exposure to the air shows that the relationship between time and
loss or gain in
moisture follows approximately the well-known equation
for slow reactions, k
log
CL
in
which a
is
the total change in
3J
at the end of
moisture content at the point of equilibrium, x the loss or gain in weight any given time t, and k the coefficient of velocity, which is a constant quantity for each kind of sugar under fixed conditions of temperature and humidity. The assumption is frequently made by samplers of sugar that the
errors
from absorption and evaporation of moisture by the sample
will
equalize one another in the long run. This, however, is far from being the case. The percentage of moisture in the ordinary grades of raw
cane sugar
is
considerably above the equilibrium point for the average

relative
humidity at the port of New York. It should be stated, howthe loss from evaporation under the prescribed conditions of that ever, is nowhere near as great as that in the above experiments, sampling
where the sugars were exposed to the open air in a thin layer. The error, however, does exist, and unless due care is exercised by the
sampler there
will be a very noticeable difference in the test. Another occasional source of error in the sampling of sugar
is
the
introduction into the sample of particles of bag, basket, mat, shavings of barrels, etc., which are introduced from the package by the trier.
The error from this cause is usually trifling; there are times, however, when it may be considerable. Such fragments of extraneous matter
do not belong to the sugar, and it devolves upon the chemist to eliminate these as far as possible before weighing out the sugar for polarizaIn removing foreign material from sample sugar the chemist tion.
must
carefully discriminate, however, between trash which belongs to the sugar and refuse which is introduced during sampling. In addition to removing trash, the chemist must complete the mix-
Lumps must be crushed and thoroughly incorporated ing of the sample. with the rest of the sample. Even samples of sugar, which are well mixed at the point of sampling, must be mixed again at the laboratory
owing to the segregation of foots at the bottom of the can or bottle. A neglect of such mixing of the sample in the laboratory is a cause of frequent differences between the results of different chemists. This mixing of the sample must be done with the utmost dispatch in order to avoid the errors due to absorption or evaporation already mentioned. Mixing of the sample upon paper or other porous substance which would absorb moisture is especially to be avoided. The method of mixing followed by the New York Sugar Trade Laboratory is as follows:
When samples are brought into the laboratory during freezing weather, the cans or bottles are first allowed to come to approximately the room temperature before opening and mixing. This is done to guard against condensation of moisture upon the cold sugar, which would lower the polarization. The sugar is poured out from the can
upon a clean sheet
etc.,
of plate glass, all pieces of bagging, baskets, mats,
thoroughly mixed with a clean steel spatula. Lumps are reduced by means of a porcelain roller and incorporated with the rest of the sample. The plate glass and porcelain roller are cleaned and wiped perfectly dry each time before using. The reduction of lumps is of greatest importance in securing uniformity of sample; the difference in polarization between the lumps and the fine portion of some sugars has been found to vary several per cent.
are removed,
and the sample
is
10
SUGAR ANALYSIS
The can from which the sugar was taken is then filled about threefourths full, the excess of sugar upon the plate being discarded. By the out of in the the little a sample can, weighing empty space leaving
by the chemist
is
facilitated.
SAMPLING OF JUICES, SIRUPS, MOLASSES, AND LIQUID SUGAR PRODUCTS

of juices, sirups, molasses, and other liquid sugar no involves special difficulties provided the material be of even products composition throughout the body of the container. A large glass or
The sampling
metal tube
may
the bungholes of hogsheads, barrels, and casks,
serve for withdrawing samples of molasses, etc., from when other means are
not available.
separately,
and
in
Containers of different capacity should be sampled making composite samples each individual fraction
it
should be proportionate to the total amount of material from which
was drawn.
The regulations of the United States Treasury Department* governing the sampling of molasses are as follows: "In drawing samples of molasses, care shall be taken to secure a fair representation and an
from each package. Packages of the be sampled in groups of not more than 25; samples from all of the packages of each group being put into a bucket. An accurate tally shall be kept and with each bucket shall be reported the number of packages the samples therein represent. The dock list accompanying the sample buckets shall convey the same information and account for every package of the mark. Packages of different size, although invoiced and permitted under the same mark, shall be sepaMolasses disrately sampled, tested, and returned for classification. charged from tank vessels shall be sampled as it is pumped from the tanks, a sample of uniform quantity being drawn at either regular
equal
of the contents
amount
same
size shall
intervals of approximately fifteen
minutes or for every 5000 gallons
discharged."
factories, various
In sampling the juices from mills and diffusion batteries in sugar automatic sampling devices have been devised for the purpose of securing a sample of the main body of juice at each instant of tune. Coomb's drip sampler (Fig. 2) is an illustration of such a device. A defect of such automatic contrivances is that they do not always give a flow of sample proportionate to the total amount of juice, f
* t
Loc.
cit.,
Art. 16.
is
J.
very efficient automatic liquid sampler Ind. Eng. Chem., 2, 253; 3, 344.
described
by G.
L. Spencer in the
11
In grinding sugar cane, when it is desired to test the work of maceration or to determine the relative efficiency of each mill, the juices from
the several sets of rollers are sampled and analyzed separately, the results of the work enabling the chemist to calculate the composition of the so-called "normal" juice or to determine the extracting power
JUICE PIPE
A.
B,
| TO | INCH VALVE. STRONG RUBBER TUBE CON-
NECTING PIPE LEADING FROM"A"WITH A GLASS T-TUBEjTO CINCHES C, INSIDE DIAMETER.
SHORT ARM OF T, FROM WHICH D, THE SAMPLE IS TO BE LED INTO AN APPROPRIATE RECEIVER.
Fig. 2.
Coomb's apparatus
for sampling sugar juices.
This phase of sampling belongs, however, to the subject and the chemist is referred to the special treatises by Spencer, Prinsen Geerligs, Deerr, and others.
of sugar-house control,
of each mill.
ERRORS OF SAMPLING DUE TO SEGREGATION OF SUGAR CRYSTALS
A serious error in the sampling of liquid sugar products is often occasioned by the crystallization and separation of sugar within the
deposition of sucrose crystals from molasses, and from and maple, cane, sorghum sirups, is an example of this; the granulation of strained honey through separation of crystallized glucose is another illustration. Containers of molasses, sirup, and honey frequently have a compact layer of crystals upon the bottom. Samples taken from the liquid surface and from the crystalline deposits of such products will show the greatest difference in composition. It is therefore necessary to mix thoroughly the contents of a container before
container.
The
sampling.
In the laboratory the crystallized sugar in a sample of sirup, molasses, or honey should be redissolved by gentle warming
12
SUGAR ANALYSIS
however, in
This is impracticable, before beginning the analysis. in bulk from casks or hogsheads, these products sampling
that the sampler can do
and the most
is to mix the contents as well as possible by and stirring. shaking The sampling of leaky containers, which allow the escape of liquid but retain all crystallized solids, is a fruitful cause of wide, and often
puzzling, discrepancies in analytical results.
ERRORS OF ANALYSIS DUE TO CHANGE

Owing
IN COMPOSITION OF
SAMPLES
to the liability of sugar products to change in composition through evaporation or absorption of moisture and through decomposition by the action of enzymes or microorganisms, it is important that
It analyses be begun as soon as possible after samples are received. in must be sent cases that for a many samples happens, however, long
distance, or stored for a considerable time, before examination can be made; the long storage of products is often necessary, as in the case of
reserve samples
which are retained
for the
purpose of confirming an
The sources of error original analysis in the event of doubt or dispute. from change in composition of samples will be briefly considered.
Changes
in
of
Absorption cause are prevented by hermetically sealing the samples in a perfectly If cans are employed all joints and connections should tight container.
Composition of Samples through Evaporation or Moisture. Changes in composition due to this
be soldered; cans of swaged metal, free from seams, are very desirable, but it has not been found possible as yet to manufacture these in large sizes. The covers should fit the cans closely and the space between
the two should be sealed
by means
of melted paraffin or
by a band
of
adhesive tape. In many respects wide-mouth glass bottles or jars are the best containers for samples; the stoppers or corks of these should
be sealed by melted paraffin or wax. In a series of experiments by Stanek
upon the drying out
of
sam-
ples of raw beet sugar in unsealed cans, the average daily evaporation of moisture for 1 month was 0.0115 per cent; when the covers of the
cans were sealed with adhesive tape (leucoplast) the average daily evaporation for 1 month was reduced to 0.0006 per cent. This loss from
evaporation is of course not evenly distributed, but is greatest during the first few days. Samples of raw cane sugar kept in covered but unsealed cans frequently show a daily increase in polarization, through
loss of moisture, of
from 0.05 to 0.10 sugar degrees during the

*
first
days
of storage.
Z. Zuckerind.
Bohmen,
34, 155.
13
Changes in Composition of Samples through Action of Enzymes. Changes in composition due to this cause are frequently noted during
the storage of plant substances, such as grains, seeds, fruits, tubers, etc. The change may consist in an inversion of sucrose by action of invertase,
in a conversion of starch
by action
of diastase, in a modification of
gums, hemicelluloses, etc., by action of other enzymes, or in a loss of It is impossible to preserve untreated sugars through respiration. plant materials of the above description for any length of time without
change in composition, although the rate of change may be greatly by cold storage. Heating the samples before storing will destroy enzymes, but has the disadvantage in some cases of causing
retarded
inversion or of liquefying
and saccharifying
starch.
Freezing the
suspend enzyme action for the time, but may on the other hand incite changes of a different character, as in the production of sucrose from starch in frozen potatoes. When samples of fresh plant materials, which are liable to undergo enzymic decomposition, cannot be analyzed immediately, an effective
material
may
method
of preventing change is to weigh out a quantity of the finely reduced substance and preserve in a stoppered jar or bottle by the
addition of alcohol.
An
excess of alcohol (over 50 per cent) destroys
the action of enzymes,
and samples thus preserved do not undergo any
change in composition after many months' standing. Changes in composition through enzyme action may also occur in It has happened in the author's experience that cold-strained honey. bottle of such a honey, which contained over 20 per cent sucrose at the time of sampling, contained after 4 months' storage less then 10 per cent; in a second sample of the same honey, which was kept in a warm laboratory during the same period, the sucrose was almost completely inverted. The inversion was probably due to an invertase secreted by the bees. The action of enzymes in such products as honey may be destroyed by heating the sample to a temperature of 80 C. Changes in Composition of Samples through Action of Microorganisms.
The
effect of yeasts,
moulds, and bacteria in changing
the composition of sugar products is well known. While the conditions for the development of microorganisms are most favorable in such dilute media as juices and musts, they may also cause deterioration in
such concentrated products as molasses and sugar. The fermentation menstruum as molasses, however, is confined entirely to the surface, which, through the attraction of hygroscopic moisture, beof such a thick
comes dilute enough to favor microorganic growth. The same is true of raw sugars; the film of molasses coating the crystals undergoes a
14
SUGAR ANALYSIS
is
gradual fermentation, with the result that the underlying sucrose

slowly dissolved
and inverted. The changes which may occur as a result of fermentation in stored samples of raw cane sugar may be seen from the following polarizations
made by Browne* at the Louisiana Sugar Experiment Station upon several samples of Cuban Centrifugal sugars after keeping 9 months in
the can.
TABLE
III
Showing Deterioration of Sugar Samples in Storage

Number.
CHAPTER
II
DETERMINATION OF MOISTURE IN SUGARS AND SUGAR PRODUCTS BY METHODS OF DRYING

accurate determination of moisture, in some respects the of analytical operations, is frequently one of the most difficult determinations which the sugar chemist is called upon to
THE
most simple
make.
Among
the chief difficulties which confront the chemist in de-
termining the moisture content of sugar products by the ordinary methods of drying, may be mentioned: (1) the very hygroscopic nature
of many sugar-containing materials and the retention of water by absorption or occlusion; (2) the extreme sensitiveness of some sugars,
notably fructose, to decomposition at temperatures between 80 and 100 C., with splitting off of water and other volatile products; (3) the liability of many impure sugar-containing substances to absorb
oxygen during drying, with formation
of acids
and other decomposition
The moisture determination is further complicated by the products. fact that many sugars, as maltose, lactose, and raffinose, retain variable amounts of water of crystallization under different conditions of drying,
so that the chemist
is
of weight occurs in the
even when no further loss not always certain as to the exact amount of moisture oven
may be retained in a hydrated form. In the following description of processes for determining moisture, methods will be given for a number of typical substances. The first
class of
which
methods to be described
to 110
is
are stable at 100
C.
The determination
intended only for products which of moisture in cane
sugar
is
taken as an
illustration.
DETERMINATION OF MOISTURE
IN
CANE SUGAR
Refined sugar, raw beet sugar, and the superior grades of raw cane sugar are dehydrated successfully by drying 2 to 5 gms. of the finely powered sample in a thin layer for 2 to 3 hours in a boiling-water oven
The in a special oven for 1 hour at 105 to 110 C. in loss the after in weight, cooled a determining sugar desiccator, and, reheated at 105 to 110 C. for another hour. The process is continued until successive heatings cause no further loss.
and then heating
is
15
16
SUGAR ANALYSIS
For weighing out the sugar flat-bottomed aluminum, nickel, or are also conplatinum dishes may be used; clipped watch glasses With lower-grade sugars, which convenient. (See Figs. 3 and 4.) tain hygroscopic salts and other impurities, the dish should be covered during weighing. For many purposes of dehydration low glass-
Fig. 3
Fig.
Fig. 5
Receptacles for drying sugar.
stoppered weighing bottles (Fig. 5) are well suited, and prevent loss of moisture in weighing out the sample and absorption of moisture in weighing the dry residue.
The
official
method*
of
the Association of Official Agricultural
Chemists for determining moisture in sugars prescribes drying in a hotwater oven for 10 hours. With some sugars, more especially those of
large grain, there is danger of occlusion and retention of water, and the The last traces of moisture may not be expelled at 98 to 100 C. method of the International Commission f upon Unification of Methods
Sugar Analysis prescribes in case of normal beet sugars drying at to 110 C.; this temperature is sufficient to expel the last traces of occluded water and is not attended with sufficient decomposition
for
105
to affect the weight of product.
The temperature
of drying
by
this
method should not exceed 110 C.

For maintaining a uniform temperature of 105 to 110 C. a glycerin may be used. The Soxhlet drying oven, shown in Fig. 6, is favored by many for rapid drying. The bath is filled with a salt solution of the desired boiling point, and closed with the condenser B. The material is placed in the oven and the door tightly clamped at A. Upon lighting a gas flame in the chimney C a
or salt-water bath
current of air
is generated through the flues at F, and, after being heated by the boiling salt solution, passes forward from the back of the drying chamber over the material to be dried. The thermometer T indicates the temperature of the drying chamber. By raising the
Bull. 107 (revised), U. S. Bureau of Chem., p. 64. t Proceedings, Paris Convention, 1900.
DETERMINATION OF MOISTURE IN SUGARS
17
Fig. 6.
Soxhlet drying oven.
Fig. 7.
Wiesnegg hot-air oven with Reichert gas
regulator.
18
SUGAR ANALYSIS
temperature gradually to 100 C. and then to 105 C. for the final dehydration, the time of -drying by the Soxhlet oven may be reduced hi many cases to less than an hour. A mixture of glycerine and water of the desired boiling point is less liable to corrode the metal of the oven
than the salt solution, and is preferred by many for this reason. In case a hot-air oven is used for drying at 105 to 110 C., the temperature should be governed by means of a gas regulator. A Wiesnegg hot-air oven with porcelain inner chamber and glass door is
a very suitable type. Illustration with Reichert gas regulator is shown In using hot-air ovens, where considerable variations in in Fig. 7. temperature are liable to occur through unequal distribution of heat,
the exact temperature of drying should be determined eter placed near the material under examination.
by a thermom-
DETERMINATION OF MOISTURE IN SIRUPS, MOLASSES, MASSECUITES, ETC., WHEN FRUCTOSE is ABSENT OR PRESENT ONLY IN TRACES
For dehydrating sirups, molasses, massecuites, and other sugarcontaining substances, which contain but little or no fructose, the method of drying previously described may be used. The material,
first be absorbed upon dry sand, pumice stone, or asbestos in order to facilitate the removal of the large excess of water.
however, should
The
following provisional methods* of the Association of Official Agricultural Chemists are recommended for drying the semiliquid products
of this class:
Drying upon Pumice Stone.

of fineness.
"Prepare pumice stone in two grades
One
of these should pass through a
1-mm.
sieve, while
the other should be composed of particles too large for a millimeter sieve, but sufficiently small to pass through a sieve having meshes
6
mm.
in diameter.
Make
the determination in
flat
metallic dishes or
in shallow, flat-bottom weighing bottles. pumice stone 3 mm. in thickness over the
Place a layer of the fine
bottom of the dish and upon this place a layer of the coarse pumice stone from 6 to 10 mm. in thickness. Dry the dish thus prepared and weigh. Dilute the sample with
a weighed portion of water in such a manner that the diluted material shall contain from 20 to 30 per cent of dry matter. Weigh into the
dish,
prepared as described above, such a quantity of the diluted sample
as will yield, approximately, 1 gm. of dry matter. Use a weighing bottle provided with a cork through which a pipette passes if this
weighing cannot be
*
made with extreme

U.
S.
rapidity.
of
Place the dish

p. 64.
in.
Bull. 107 (revised),
Bureau
Chem.,
19
a water oven and dry to constant weight at the temperature of boilIn case of ing water, making trial weighings at intervals of 2 hours.
materials containing
much
levulose or other readily decomposable
substances, conduct the drying in vacuo at about 70 C." " In a flat-bottom dish place 6 to 7 gms. Drying upon Quartz Sand. short sand and a of pure quartz stirring rod. Dry thoroughly, cool in
a desiccator, and weigh. Then add 3 or 4 gms. of the molasses, mix with the sand, and dry at the temperature of boiling water for from
8 to 10 hours.
Stir at intervals of
and weigh. Stir, Repeat heating and weighing weigh. is not greater than 3 mgs.
"
an hour, then cool in a desiccator, heat again in the water oven for an hour, cool, and
until loss of
water in one hour
Before using, digest the pure quartz sand with strong hydrochloric acid, wash, dry, ignite, and keep in a stoppered bottle." In order to prevent the occlusion or retention of water in the dried
residue, an hour of drying at 105 to 110 the determination of moisture in sugar.
C.
is
advisable as under
In a method of Determining Moisture.* Pellet nickel in France, capsules, 85 mm. drying considerably employed
Pellet's of
Method
3
Fie;.
Fig. 9
Pellet capsule for drying liquid sugar products.
wide and 20
mm.
deep, are used.
in the center as
shown
in Fig.
The capsule has a circular depression Each capsule is provided with a 8.
cover having a small notch at the edge for the passage of a small stirring
rod.
The
1
raised border of the capsule
is filled
with
fine particles (about
diameter) of freshly ignited pumice stone, employing an inverted The funnel is then removed, the cover and funnel as shown in Fig. 9.
mm.
rod put in place, and the capsule weighed. Three grams of the substance to be dried are then weighed in the central depression of the capsule; 5 c.c. of hot distilled water are then added, and after
stirring
*
"
Fribourg's
Analyse .chimique" (1907), pp. 90-94.
20
SUGAR ANALYSIS
different sides to permit absorption of the solution The process is repeated with 3 c.c. more of hot
is slightly inclined stirring to dissolve all soluble matter, the capsule
on
by the pumice
water and then with 2 c.c. The contents of the capsule are then spread evenly over the entire bottom and dried in any suitable oven at a final temperature
stone. of 102
to 105
C.
In case of products containing even traces of free acid, a drop or two of strong ammonia is added. The excess of ammonia is expelled and the amount retained in the combined form is usually too small
to be regarded.
If
the free acid
is
not neutralized, inversion of sucrose
may
result,
with the introduction of a considerable error in the deter-
mination.
DETERMINATION OF MOISTURE IN PRODUCTS WHICH CONTAIN FRUCTOSE

Owing to the susceptibility of fructose to decomposition in presence water at temperatures much above 70 C., the methods previously described are not applicable to the determination of moisture in such
of
products as honey, sugar-cane molasses, jams, fruit products, and other similar substances. The error which may result from this source
may
be seen from the following experiment by Carr and Sanborn upon dehydrating a solution containing 17.75 per cent of fructose. The
solution
in air.
was dried upon pumice stone

Hours
of drying.
in flat-bottomed dishes at 100
C.
21
atmospheric pressure is the only recognized method for the accurate determination of moisture in fructose-containing materials. Carr and Sanborn's Method. Many methods have been devised
The following process is the for drying sugar solutions in vacuum. one described by Carr and Sanborn,* who have employed their method successfully upon the widest range of materials, such as fructose solutions,
honey, molasses, sorghum and maize
juices, etc.
Select clean, fine-grained pumice stone and divide into fragments the size of No. 4 shot. Pass the dust through a 40-mesh sieve and
"
from the larger particles. Digest hot with 2 per cent sulphuric acid and wash until the last trace of acid disappears from the wash water. Owing to the ready subsidence of the material, the washtreat separately
may be accomplished rapidly by decantation. After complete washing, place the material, wet, in a Hessian crucible, and bring to redness in a monitor or other convenient furnace. When complete expulsion of water is assured, place, hot, in a desiccator, or direct into
ing
the drying dishes if desired for use immediately. In loading the dishes place a thin layer of the dust over the bottom of the dish to prevent
contact of the material to be dried with the metal; over this layer place the larger particles, nearly filling the dish. If the stone has been well washed with the acid, no harm may result from placing the dish
and stone over the flame
for a
moment
is
before placing in the desiccator
preparatory to weighing. " If the material to be dried

is
dense, dilute until the specific gravity
neighborhood of 1.08 by dissolving a weighed quantity in a weighed quantity of water. (Alcohol may be substituted in material not precipitable thereby.) Of this, 2 to 3 gms. may be distributed over the stone in a dish, the area of which is in the neighborhood of
in the
3 sq.
in.,
or
gm.
for each square inch of area.
Distribute this material
uniformly over the stone by means of a pipette weighing bottle (weighing direct upon the stone will not answer), ascertaining the weight taken by difference. " Place the dishes in a vacuum oven, in which may be maintained a
pressure of not more than 5 in. mercury, absolute. The form of oven is not material so long as the moisture escapes freely by passing a slow current of air (dried) beneath the shelf supporting the dishes.
The temperature must be maintained

25
in.
at 70
C. and the
vacuum
at
must be taken when the dish is covered by a ground the plate, and open dish must not be exposed to the air longer than * Bull. 47, U. S. Bureau of Chem., pp. 134-151.
All weighings
"
22
absolutely necessary. 3 hours."
SUGAR ANALYSIS
Weighings should be made at intervals of 2 or
The following triplicate series of experiments were made by Carr and Sanborn upon a solution containing 17.10 per cent fructose. The solution was dried on pumice stone in flat-bottomed dishes at 70 C.
under a vacuum of 25
Hours.
in.

of thermometer,
23
and for inlet and exit of air. A gas drier containconcentrated sulphuric acid may be used for removing moisture ing from the slow current of entering air. The detachable plate at the end of the oven is provided with a rubber gasket and is fastened into
position
by four screws which secure a
Browne's Method of Vacuum Drying. constructed types of vacuum drying oven
perfectly air-tight joint. When one of the specially

is
not available, the author
has found the following arrangement (Fig. 11), which is easily constructed from ordinary laboratory materials, to be perfectly efficient.
I
->To Vacuum Pump
Fig. 11.
Browne's method of vacuum drying.

consists of a large-mouth bottle (B) of
The vacuum chamber

glass,
heavy
supported by the shelf (S) of an ordinary water oven The mouth of the bottle is closed by a tight-fitting rubber stopper (0). of the (R) whose 3 holes permit the insertion, through the top opening is bottle The T. thermometer oven, of the tubes I and E and the
which
is
easily fitted,
shelf,
and detached from the stopper by
first
withdrawing the
the bottle is in the latter being shoved into position bottom of the the / to tube The current of air entering by place.
again when
24
SUGAR ANALYSIS
vacuum bottle is controlled by a clamp pinchcock (C) and freed of moisture by a gas drier (D). The exit air from the vacuum bottle passes by the tube E to the vacuum pump or aspirator.
For absorbing the sugar-containing
liquid, asbestos
in perforated
The tubes measure 9 cm. long by 2 cm. brass or copper tubes is used. in diameter, and are nearly filled with freshly ignited asbestos, the latter being tightly packed with a rod against the sides in the upper
half of the tube, so as to leave a central cavity. Each tube thus prepared is placed in a glass-stoppered weighing About 5 c.c. of the bottle of sufficient size, and the whole weighed.
liquid to be analyzed are then delivered from a pipette into the cavity in the asbestos, the object of the cavity being to secure a rapid absorption and even distribution of the liquid through the asbestos.
then immediately stoppered and reweighed, the amount of substance taken. After removing the stopper the weighing bottle with tube is placed in the in the diagram, and the temperature vacuum bottle, as shown by raised to 70 C. During the first few hours of drying a brisk current
bottle
is
The weighing
increase in weight being the
of air is
drawn through the vacuum
bottle in order to
remove the
large excess of moisture first given off.
In the
last stages of the dry-
At the end
ing the air current is decreased and the vacuum kept at about 25 in. of a few hours the weighing bottle is removed, allowed to
cool in a desiccator,
and then restoppered and weighed. The bottle is then redried for a second short period to determine if all moisture has been expelled. In the weighing out of juices, sirups, sugar solutions, etc., for absorption
upon pumice stone, sand, or asbestos, a small flask provided with a stopper and a rubber-bulbed pipette or medicine dropper
will
be found convenient (Fig. 12). The bottle is about two-thirds full with the sugar solution, which should not contain over 25 per cent solids,
filled
Fig.
12.
Bottle for
~
and then closed with the stopper and pipette. After weighing the bottle and contents, about 5 c.c. f Uquid are conve y ed b y means of the bulb Pêtte to the absorbent material, and the flask restoppered
difference in weight
and weighed.
is the amount of sample taken. Honeys, molasses, jellies, and other water-soluble substances of high density should be diluted before this method is employed, by dissolving a weighed amount of substance in a weighed amount of water.
The
The above method
of
weighing samples
is
precluded, however,
when

insoluble matter
is
25
present, as with jams, sauces, and similar products. In such cases a weighed amount of the well-mixed sample is stirred
with a
little
water until
all
soluble matter
is
dissolved
and then com-
pletely transferred to the absorbent material in the drying dish with help of a fine jet of water. The Pellet method of drying is especially
convenient for products of this
class.
DETERMINATION OF MOISTURE IN SUGAR MATERIALS WHICH CONTAIN WATER OF HYDRATION

Difficulty
is
sometimes experienced in dehydrating sugars such as
glucose, lactose, maltose, and raffinose, which crystallize with one or more molecules of water of crystallization. The principal precaution
to be observed in drying such sugars is not to raise the temperature in the first stages of the process above the melting point of the hydrate,
otherwise the sugar will liquefy to a thick viscous mass from which it is difficult to expel the last traces of water without decomposition. For drying glucose hydrate, C 6 Hi2 O 6 2 0, the sugar is spread in
+H
a thin layer and gently warmed at 50 to 60 C. for several hours, when most of the water will be removed without melting of the crystals.
The sugar
is
then gradually heated to about 105
C.,
when
the last
traces of water will be expelled, with no evidence of liquefaction. 5 For drying raffinose hydrate, Ci 8 32 Oi6 2 O, the finely powdered
+ H
sugar is first warmed to 80 C. for several hours and then the temperature gradually raised to about 105 C. The preliminary drying may be hastened greatly by heating the sugar in a vacuum oven.
H 2 O, gives off its water very incomMaltose hydrate, Ci 2 H 22 On under atmospheric pressure, and vacuum dehydraThe sugar is gently heated under a strong vacuum at 90 to 95 C., and then after a few hours the temperature is raised to between 100 and 105 C. H2 O, retains its water of crystallizaLactose hydrate, Ci 2 H 22 On
pletely at 100 C. tion is necessary.
tion
unchanged at 100 C. under atmospheric pressure. It is therefore customary in analytical work to estimate lactose as the hydrate. Lactose may be dehydrated, however, by gently heating the finely pulverized sugar in a strong vacuum to a temperature of 125 to 130 C. The method of drying devised by Lobry de Bruyn and van Laent,* and used Morris, and Millar,f and also by Walker,t is to
by Brown,
*
weigh the finely powdered sugar in a small flask and connect the latter
Rec. trav. chim. Pays-Bas, 13, 218.
t J. j J.
Chem.
Am. Chem.
Soc. Trans., 71, 76. Soc., 29, 541.
26
SUGAR ANALYSIS
tube to a bottle containing phosphorus pentoxide, P2 5 as a dehydrating agent. The open branch of the T tube is connected with
by a
a strong vacuum; the flask containing the sugar is then placed in an oil bath and the temperature gently raised to the point desired. Walker found that lactose under these conditions, after heating 1 hour at 80 C. and then 1 hour at 130 C., remained perfectly white, but upon heating to 140 C. the sugar became tinged with brown, showing signs of decomposition.
The method
cessfully
Lobry de Bruyn and van Laent has also been suc* employed by Rolfe and Faxon for determining the total carof
bohydrates in acid-hydrolyzed starch products. In the modified apparatus of Rolfe and Faxon the T tube is provided with a three-way
stop-cock, which allows the great excess of water first given off to be removed without coming in contact with the phosphorus pentoxide.
*
J.
Am. Chem.
Soc., 19, 698.
CHAPTER
THE
of
III

quantity of matter in a unit volume of substance is called the If be the mass and V the volume
absolute density of that substance.
a given substance,
its
absolute density
will
be
D= =
The
between the masses of equal volumes of a substance and of some standard material is the relative density of that substance. Since, however, the masses of two bodies at any one place are proportional to their weights, the relative density S of a given substance may be exratio
pressed
w S = ^> where w and
W are the weights respectively of
equal
is
volumes of the substance and standard material.
Relative density
commonly known
of
as specific gravity, and, since the standard substance
comparison
is
is
defined as a
tion
number
nearly always water, specific gravity is commonly indicating how much heavier a substance or soluof water.
is
than an equal volume
The determination
of specific gravity
one of greatest importance
in the analysis of sugars; its great value consists in the fact that solu-
tions of different sugars of equal concentration
have very nearly the
same
to
following specific gravities are given for specific gravity. 10 per cent solutions of nine different sugars at 20 C. with reference
The
Arabinose 1.0379, glucose 1.0381, fructose 1.0385, sorbose 1.0381, sucrose 1.0381, maltose 1.0386, lactose galactose 1.0379, It will be noted that the specific gravity of raffinose 1.0375. 1.0376,
water at
4C.:
each sugar solution is but little removed from the average 1.0380, which It is possible, therefore, by means is almost the same as that of sucrose.
of specific gravity tables established for solutions of pure sucrose to determine very closely the percentage of dissolved substance for any
sugar or mixture of sugars in aqueous solution. Units of Volume. The unit of volume universally employed in sugar analysis is the cubic centimeter. This unit is differently defined and the chemist must distinguish carefully between (1) the metric or
true
cubic centimeter, (2) the reputed cubic centimeter.
Mohr
27
cubic centimeter, and
(3)
the
28
SUGAR ANALYSIS
The Metric Cubic Centimeter is defined as the volume occupied by one gram of water weighed in vacuo at 4 C., the temperature of maximum density (D = 1.000000). At 20 C. the metric or true cubic centimeter is equivalent to the volume occupied by 0.998234 gram of water weighed in vacuo, or 0.997174 gram of water weighed in air with brass
weights.
gram
The Mohr Cubic Centimeter is defined as the volume occupied by one One Mohr of water weighed in air with brass weights at 17.5 C.
is
cubic centimeter, as thus defined, centimeters.
equivalent to 1.00234 metric cubic
The Reputed Cubic Centimeter, a term introduced by Brown, Morris, and Millar,* is defined as the volume at 15.5 C. of one gram of water weighed in air with brass weights. One reputed cubic centimeter, as
thus defined, is equivalent to 1.00198 metric cubic centimeters. The true or metric cubic centimeter was adopted as the standard
unit of volume
of
by the International Commission
for
Uniform Methods
Sugar Analysis at its meeting in Paris, 1900.

SPECIFIC GRAVITY TABLES FOR SUGAR SOLUTIONS
Various tables have been established by different observers which

give the specific gravity (sp. gr.) of cane-sugar solutions for different These tables are expressed in several ways; they vary concentrations. according to the temperature which is selected for the determination,
15 C., 17.5 C., or 20 C. being usually taken, and also as to whether the weight of water at 4 C. (true specific gravity) is used for comparison, or water at 15C., 17.5C., and 20 C. (relative specific gravity). In expressing specific gravity it is customary to indicate the system employed by writing the temperature of the solution above that of the
-
water; thus,
^> ^ jf f, etc. In Table IV the specific gravities

,
.
of sucrose solutions at several

different
concentrations are given according to the calculations of

authorities.
Various formulae have been worked out for expressing the relationship between the specific gravity and percentage by weight of dissolved
sucrose.
Gerlach for specific gravity

the equation
]! has
expressed the relation-
ship
by
y
= 1+ 0.00386571327 Z
is
+ 0.00001414091906 z + 0.0000000328794657176 z
2
3
,
in
which y
the specific gravity and x the per cent of sugar.

*
J.
Chem.
Soc., 71, 78 (1897).

of different temperatures with the following results:
Temperature.
29
Scheibler has recalculated Gerlach's equation for sugar solutions
10
15
20 30
40
50 60
y=l + 0. 003976844 x + 003915138 x + y = 1 + y = l + 0. 003884496 x + = 1 + 0. 003844136 x + = 1+0. 003796428 x + = 1 + 0. 003764028 x + 0=1 + 0. 003722992 x + = 1 + 0. 0036831 12 x +
.
0000142764 x 2 0000139524 x 2 0000139399 z 2 0000144092 x 2 0000145456 x 2 0000143700 x 2
0000148088 x 2 0000155904 x 2
+ + + + + + + +
000000029120 x 3 000000032728 z 3
000000033806 z 3 000000030912 x 3 000000030664 x 3 000000035192 x3
000000032440 x 3
000000026368 x 3
TABLE IV
Specific Gravity of Sucrose Solutions by Different Authorities
Sucrose, per cent
by weight.
30
SUGAR ANALYSIS
most recent and most accurately established tables are those of the German Imperial Commission* upon Standards, based upon the determinations of Plato, and published in 1898 and 1900. These tables
give the percentages of sucrose for specific gravities at y^'
15*
>
and
^r.
The ^?
table,
which was established according to the require-
ments
of the
Fourth International Congress of Applied Chemistry
given in the Appendix (Table 1). The specific gravity tables of the German Imperial Commission have since been enlarged by Sidersky,f so as to give the grams of sugar
(Paris, 1900), is
between 100 gms., and also for 100 c.c., of solution for ^ and Brix. 30 concentrations between and For for 10 and 30 C. and their limited range Sidersky's tables are the most complete of any
for
which have been compiled.

Influence of Temperature upon the Specific Gravity of Sugar
Solutions.
With
increase of temperature, sugar solutions
expand
less.
in
volume and the

concentration.
specific
gravity becomes correspondingly
The
coefficient of cubical
Josse and
expansion of sugar solutions varies according to RemyJ give the following coefficients for
different sugar solutions
between 15
and 25
C.:
TABLE V
Coefficients of Cubical
Expansion for Sugar Solutions
d!5C.

Knowing the value
equation
dt
31
can be calculated from the
of 7, the specific gravity dt at temperature t specific gravity dt at temperature to by the
methods
In the employment of temperature corrections in densimetric of analysis, it is more customary to apply the correction to the
percentage of sugar (degrees Brix) rather than to the specific gravity. The correction is to be added in case the temperature is above, and
to be subtracted in case the temperature is below, the standard degree of the table (17.5 C. for the old Brix tables and 20 C. for the new
tables of the
German Commission).
Lists of such corrections are
affixed to the standard tables of specific gravities.* Determination of Dissolved Solids by Use of Solution Factors.
In the investigation of starch-conversion products the percentage of solids in 100 c.c. of solution is frequently calculated from the specific " gravity by means of a solution factor." This method was introduced
in 1876
by O'Sullivan, f who found
that,
when
10 gms. of maltose or
c.c.,
dextrin were dissolved at 60 F. (15.5
C.) to 100
a solution of
1.0385 sp. gr. (jf^) was obtained. Assuming that the percentage of dissolved substance is always proportional to the specific gravity of the solution (which is only approximately true), a solution containing 1
gm. of maltose or dextrin in 100

of 1.00385 at 15.5
c.c.
should have a specific gravity
C.
solution of specific gravity d should contain

,
at
KKOr 15.5 C.
,
1000 -
(d- 1.000) o.oO
gms. of
solids.
Brown, Morris, and Millar J determined the solution
factors of a
{575-0
number of different sugars for a uniform with the following results:

TABLE VI
specific gravity of 1.055
Solution Factors of Sugars and Starch Conversions
Anhydrous Anhydrous Anhydrous Anhydrous Anhydrous
glucose ..................................... sucrose ....................................

'
3 825
.
3 859
invert sugar ................................ fructose ....................................
3 866
.
3 907
.
maltose .................................... Low starch conversion ([]/> = +149.7) .......... ...... Medium starch conversion ([a] D = +173.9) .............. High starch conversion ([a] D = +188.6) ................ Dextrin ................................................
-
3 916
.
3.947 3.985 4.000

4.206
Appendix, Tables 2 and

J J.
4.
t J-
Chem.
Soc. (1876), 129.
Chem.
Soc. (1897), 71, 72.
32
SUGAR ANALYSIS
The
solution factors of glucose, fructose,
*
been determined by Ling, Eynon, and Lane

results as
and maltose have recently with practically the same
Brown, Morris, and Millar. For ordinary purposes Brown, Morris, and Millar recommend the use of the sucrose factor 3.86. A comparison of the actual grams of sucrose per 100 c.c. of solution with those calculated by means of this
solution factor
is
given in the following table:
TABLE VII.
15.5
'
15T

nomenon
of
33
contraction in volume during solution of sucrose and water has long been known. It was first observed by Reaumur and Petit le Medecin in 1733, and has been repeatedly studied by many
subsequent observers.* ously estimated. If x

in
volume
extent of this contraction has been varithe per cent of dissolved sucrose, the change according to Brixf is represented by the equation
is
The
0.0288747 x
0.000083613 z 2
0.0000020513
x*.
Scheibler J gives the equation
0.0273731 x
0.000114939
x*
0.00000158792 x 3
according to which the maximum contraction is 0.8937 c.c. for 55.42 gms. sucrose and 44.58 gms. water at 17.5 C. Gerlach gives the maximum contraction as 0.9946 c.c. for 56.25 gms. sucrose and 43.75 gms.
water,
others, the maximum contraction is reached at about 40 per cent sucrose; beyond this there is a decrease until at 60 per cent sucrose the contraction is 0; with concentrations
as 0.9958 Ziegler to Matthiessen According
and
c.c. for
56 gms. sucrose and 44 gms. water.

||
and
above 60 per cent sucrose there

of the question
is
is
an expansion
in volume.
This view
due, according to Plato, 1f to the mistaken idea that dissolved sucrose has the same specific gravity as the crystallized solid
(1. 59103
|p
for chemically pure
powdered
sucrose, 1.5892 1
for chemi-
cally pure sucrose crystals).
If
we take
Plato's calculated value for
the specific gravity of dissolved sucrose in aqueous solution, 1.55626, the following results (Table VIII) are obtained which are in close concord-
ance with those of Gerlach and Ziegler. The apparent change in specific gravity of dissolved sucrose is due to the phenomenon of contraction, for which no satisfactory explanation has as yet been offered.
*
In contradiction to the results of
all
previous experimenters, Olizy (Bull.

experiof sucrose in
assoc. chim. sucr. dist., 27, 60) claims to
have demonstrated by numerous
ments that absolutely no contraction takes place during the solution

water.
t Z.
Ver. Deut. Zuckerind., 4, 308.
Neue
Zeitschrift, 26, 37. Oest. Ung. Z. Zuckerind., 12, 760.
II
Lippmann, "Chemie der Zuckerarten," 1081.

Z. Ver. Deut. Zuckerind., 50, 1098.
If
34
SUGAR ANALYSIS
TABLE VIII
Showing Contraction in Volume of SucroseWater Mixtures
Per cent
sucrose.

The
35
While the Specific Gravity of Impure Sugar Solutions. application of specific gravity tables established for sucrose to the estimation of dissolved substance in solutions of other sugars and carbohydrates is fairly accurate, their use in the case of impure sugar solutions may lead to serious errors, owing to the fact that the percentage of dissolved impurities for the same specific gravity differs from the corresponding percentage of sucrose. The errors resulting
from
this cause
may
be seen in Table X, which gives the concentrations
of sucrose, tartaric acid,
sodium potassium tartrate, and potassium carbonate for different specific gravities. When the specific gravity is determined after dilution with a definite amount of water, as is necessary with very thick sirups, the error in estimation of dissolved substance
is still
further intensified, owing to the difference in contraction
TABLE
X
Com-
Concentrations of Aqueous Solutions of Organic and Inorganic pounds Compared with Those of Sucrose at 15 C. for
the
Same
Specific Gravity
Specific gravity.
36
SUGAR ANALYSIS
between sugar and dissolved impurities in aqueous solution. This can be seen by reference to Table X; it is also shown in Table XI, which
gives the calculated differences in contraction obtained by diluting solutions of sucrose, tartaric acid, sodium potassium tartrate, and potassium carbonate with water to reduce degrees Brix from 50 to 10.
differences
Additional comparisons showing the between true dry substance

as calculated from
and dry substance
specific gravity are given for a of compounds in Table XVII.
number
METHODS OF DETERMINING
SPECIFIC
GRAVITY OF SUGAR SOLUTIONS
by means
In the estimation of dissolved sugars of specific gravity, the tem-
perature of the laboratory is not always the same as that prescribed by the table. It is then necessary either to bring the
solution to the required temperature by artificial means or else to apply a fixed
correction from a conversion table.

latter
The
method
is
the more convenient
and
for ordinary purposes is sufficiently
in cases, however, where great accuracy is required the determination must be conducted under absolutely the
exact;
same temperature conditions

fied in
as speci-
the tables.
Specific Gravity Bottle or

eter.
PycnomThe most accurate method for
Fig.
13.
Specific gravity bottle equal with thermometer. tion.
the determination of specific gravity is the direct comparison of the weights of
volume
is
of water and sugar soluIn this method some form of
specific gravity bottle or
pycnometer
used, various types of which
are
shown
in Figs. 13 to 16.
Before using the instrument the pycnometer is calibrated by determining the weight of distilled water which it contains at the temperature of comparison. The bottle is first thoroughly cleaned by means of dilute caustic soda and hydrochloric acid; it is then washed with distilled water and dried in an air bath. In case of pycnometers

warmed beyond the
37
constructed with a thermometer stem, the latter should never be

limit of graduation, which is frequently only 40 C., otherwise the expansion of the mercury may break the instrument. After drying and cooling the pycnometer is weighed. The bottle is next filled with distilled water, recently boiled and cooled to
The temperature adjustment is best effected by expel dissolved air. water a degree or so lower than the temperature with the bottle filling is then inserted, taking care to prevent the introthe stopper desired;
duction of air bubbles, and the bottle placed in a bath of water kept After about 10 minutes, or as exactly at the desired temperature.
Fig. 14
Fig. 15
Fig. 16
Types
of specific gravity bottles.
soon as the thermometer of the instrument has risen to the right degree, the excess of water,
wiped perfectly dry and reweighed. The increase in weight is the water capacity of the bottle at the desired temperature. The process is repeated and the average of several determinations used
as a constant in
to be examined,
all
tion mark, is and the bottle
exuding from the stem, or above the graduaremoved with a thin piece of filter paper, the cap is fitted,
subsequent work.
after redrying or rinsing repeatedly with the liquid next filled with the sugar solution (observing the
The pycnometer,
is
same precautions
specific gravity.
as to temperature as before)
weight of solution divided by
and reweighed. The the water capacity of the bottle gives the
of
Since 20 C. has been adopted as the standard temperature* for At the sixth session of the International Commission for Uniform Methods to accept Sugar Analysis (London, May 31, 1909) it was "voted unanimously
*
a single specific gravity table as standard, at the temperature of 20 C., which is to be based upon the official German table. From this, other tables may be calculated at other temperatures, for instance, at 15 C., 17.5 C., 30 C., etc."
38
all
SUGAR ANALYSIS
it is
processes of sugar analysis,
best to
make
the determination of
specific gravity
when
possible at this temperature.

|j-
For the
specific
gravity
at 20
the value for
must be multiplied by the density
of water
C., or 0.998234.
For very exact work the calculation of specific gravity must be made upon the weights in vacuo, in which case a correction for the density of the air must be introduced. The method of making the calculation is as follows Let A = apparent weight of pycnometer, B = ap= apparent weight parent weight of pycnometer and water at t C., C of pycnometer and sugar solution at t C., d = density of water at = density of air at t C. and the observed atmospheric t C., and s the corrected specific gravity S will be then pressure;
:
C~A
B~C
If the temperature of the laboratory is much above that of adjustment, the specific gravity bottle and contents must remain at rest until they acquire the surrounding atmospheric temperature, otherwise moisture will condense upon the instrument and interfere with the weighing. It is needless to add that the cap of the bottle must be suffi-
ciently tight to prevent leakage of liquid displaced by expansion through increase of temperature. Pycnometers whose stems are to be filled
to
mark and hence allow room

be preferred.
for expansion, as Fig. 13, are gener-
ally to
For certain kinds
of
work
(as for densities of
very dilute sugar solutions) Sidersky* recommends Boot's pycnometer (Fig. 15), which, having a double wall with vacuum, keeps the temperature of the solution constant for a long time.
For highly concentrated sugar

cuites,
solutions, such as molasses, masse-
or other viscous substances, the

if
method must be somewhat
modified,
the specific gravity of the undiluted material is desired. For this purpose a pycnometer with rather wide neck, of the form in
Fig. 16, is chosen, and filled nearly to the mark with the hot material to be examined. To remove occluded air bubbles the bottle is placed
for a short
is
or salt-water bath, the boiling point of which keep the material in a liquid condition. After cooling to 20 C. and weighing, the space between the substance and the graduation mark is filled with distilled water and the bottle reoil
time in an
sufficiently high to
weighed.
The method of example upon a molasses:

* "
calculation
is
illustrated
by the following
Les Densit^s des Solutions sucrees," p.
17.

A, water capacity of pycnometer B, weight of molasses C, weight of molasses and water C B = weight of water added A (C B) = weight of water
occupying space of molasses
56.348
39
= = =
=
50.124 gms. 56.348 gms. 66.536 gms.

10.188 gms.
39.936 gms.
39.936
1.411 sp. gr. of molasses.
to
mark
Reich* has modified the above method by filling the pycnometer directly from a burette divided into 0.05 c.c. and noting the
Fig. 17.
Determining
specific gravity
by means
of analytical balance.
volume
as the top of water added. If the burette has 50 instead of graduation, the actual cubic centimeters of molasses, etc., in the pycnometer is read off directly when the latter is calibrated to hold exactly 50 c.c. This of course obviates a second weighing of the pycnometer,
and, while not as accurate as the method of weighing,

for
is
sufficiently close
many
purposes:
solutions
second method for determining the specific gravity of sugar that is based upon the well-known principle of Archimedes,
*
Deut. Zuckerind., 34, 38.
40
SUGAR ANALYSIS
a body immersed in a liquid loses the same weight as that of the volume It is therefore only necessary to compare the of liquid displaced. losses in weight which the same body undergoes in water and in a given
process
is
The solution, in order to determine the specific gravity of the latter. may be carried out in a variety of ways; a common method
by means
of the analytical balance.
A
and
sinker of
heavy
glass, or
attached to a
is
silk
thread and weighed

sugar solution.
is
a bulb of glass containing mercury, is first in air, then in distilled water,
finally in the
The method
of conducting the
weighing
shown in Fig. 17. The method of calculation
shown by the following example:

at 20 at 20
A, weight of sinker in air B, weight of sinker in water C, weight of sinker in sugar solution,
= 25.345 gms. = 22.302 gms. = 21.504 gms. == Specific gravity of sugar solution, S
C.
C. C.
1.2622
at 20
^^
s,
~
=
den-
To
convert to true density with reference to weights in vacuo, the
above equation becomes S $>

sity of
(d
s) -:
t
in
which d
water at t, and
density of air at
and the observed atmos-
pheric pressure.
Mohr's
of
Mohr, Westphal balance, makes use of the principle of the sinker described in the previous section. The construction and operation of the balance The beam (AC) of the balance is are best understood from Fig. 18. B the between at and pivoted pivot and point of suspension (C) is divided by notches into 10 equal parts. The distance between each division of the beam is ordinarily made exactly 1 cm. The balance, as usually supplied, has a specially constructed thermometer sinker (Reimann's thermometer body) which by careful grinding of the lower end is made to displace exactly 5 gms. of distilled water at 15 C.
The specific gravity balance Specific Gravity Balance. as improved by Westphal, and hence frequently called the
The
is attached by means of a fine platinum wire to the brass the combined weight of sinker, wire, and hanger being made hanger H, to equal exactly 15 gms. Before using, the balance is first adjusted
sinker
by hanging the sinker from the arm and regulating the screw S until, when the beam is at rest, the pointers of the arm and support at A
If the sinker be now submerged in distilled water exactly coincide. at 15 C., it will require 5 gms. at the point of suspension C to re-
store equilibrium.
The standard weight
for
Reimann's thermometer
41
body is therefore 5 gms., and in determining the specific gravity of solutions heavier than water this weight must always be hung from
the point C. To obtain the decimal figures of the specific gravity, weights are added to the notches on the beam until the pointers indicate The first decimal figure is obtained by means of a dupequilibrium.
licate
5-gm. weight, which
is
moved from notch
to notch
on the beam
Fig. 18.
Mohr's
specific gravity
balance (indicating 1.1267 sp.
gr.).
until the correct decimal
is
secured;
the second decimal figure
is
ob-
tained
by means
of a 0.5-gm. weight, the third decimal figure
by a
The
0.05-gm. weight, and the fourth decimal figure by a 0.005-gm. weight. beam specific gravity is then read from the scale divisions of the
in the order of the diminishing weights.

easily
The method
of reading
is
understood from Fig. 19. In using the Westphal balance the temperature of the solution is read from the thermometer of the sinker. In case of turbid or dark-
42
SUGAR ANALYSIS
colored solutions which render the reading of this thermometer difficult

or impossible, the temperature is read either by carefully drawing up the thermometer body until the top of the mercury column is visible,
by means of a larger thermometer immersed in the solution. Thermometers and cylinders of special form have been constructed for taking specific gravities, a type of which is shown in Fig. 20.
or, better,
0.9570
1.2646
1.4826
Fig. 19.
Method
of reading Westphal balance.
Fig. 20.
mometer
Special cylinder and therfor Westphal balance.
ity of
third method of determining the specific gravsugar solutions, and the one most commonly employed in technical In its usual form (Fig. 21), operations, is by means of the hydrometer.
Hydrometers.
instrument consists of a hollow glass body terminating at its lower extremity in a bulb (which can be weighted with mercury or shot) and at its upper extremity in a hollow slender stem, inside of which
this
If this instrument is allowed to float in a a paper scale is sealed. the of solution, weight liquid displaced is equal to the weight of the
43
If placed in solutions of different concentration, floating hydrometer. the stem will sink to varying depths; that point upon the scale which
is
level
for the given concentration
with the surface of the liquid indicates the density or percentage and temperature. It is in this
are calibrated
manner that hydrometers
and standardized.
In actual practice a hydrometer scale is standardized at only a few of its points, the intermediary divisions being determined by interpolation. The method of interpolation will depend upon whether the scale is to indicate
specific gravity or direct percentages.
The
weight
specific gravity
of a solution
is
equal to the
of the
hydrometer divided by the volume
of
the part submerged.
Then
W
-~
If
the scale
is
to be
graduated for
specific gravity the
numerical divisions will
proceed in arithmetical progression, The difference 1.10; 1.15; 1.20, etc.

of the
such as 1.00; 1.05;
between the volumes
hydrometer
for
any two
scale divisions will give
the volume v between these divisions; letting r
half the
diameter of the stem, then
=
^
the distance between the
two
divisions.
sions of a
relationship between the stem divihydrometer weighing 20 gms. and with a cross
2
The
area of stem
(irr )
:
equal to 0.2 sq. cm. can be seen from
the following table
TABLE XII
Showing Hydrometer Scale Divided According
Specific gravity
to Specific
Gravity
(D).
44
SUGAR ANALYSIS
It will be noted that as the specific gravity increases the distance between the scale divisions decreases. Owing to the great labor involved in the making of calculations and measurements, the division
of a
hydrometer scale harmonically
is
accomplished in practice by means
of a dividing engine.
In the graduation of a hydrometer scale for indicating direct percentages of sugar, the distance between the scale divisions is much more uniform. The relationship is best seen from the following table, where
a hydrometer of 20 gm. weight and was used as before.
0.2 sq. cm. cross area of
stem
(wr
2
)
TABLE XIII
Showing Hydrometer Scale Divided According
to
Sugar Percentage
Percentage sugar.

work.
45
the two scales are so slight that they have no significance in practical
degrees, 0-10, 10-20, 20-30, 30-40, etc., graduated into 0.1 degree. For greater accuracy a third form of spindle has been made with a range of only 5 degrees, 0-5, 5-10, 10-15, 15-20, etc., and graduated into 0.05 degree. With the help of a spindle for only approximate work, the choice of
The Brix hydrometer* or spindle is supplied in a variety of forms. For approximate work spindles are used with graduation of 0-30, 30-60, and 60^90, and divided either into 0.5 or 0.2 degree. The forms in most common use, however, have only a range of 10
Fig. 22.
Floating Brix
spindle.
Fig. 23.
Winter's cylinder for taking

specific gravity.
the particular hydrometer for the finer reading will be facilitated. The accuracy of the spindle is of course the greater, the smaller the
diameter of the stem and the consequently larger interval between the
scale divisions.
In determining specific gravity by means of the hydrometer, a tall, narrow cylinder is usually employed for holding the liquid to be examined.
*
The
spindle
is
carefully lowered into the solution in such a
The term saccharometer, which is sometimes applied to a hydrometer indicating percentages of sucrose, is unfortunate, owing to the confusion with the word saccharimeter, of entirely different meaning.
46
SUGAR ANALYSIS
way
that the surface of the stem above the liquid is not moistened. Care should also be exercised that the instrument floats freely and does
not touch the bottom or walls of the cylinder. The is the a made level with by bringing eye upon reading
BRIX.
10
the surface of the solution and noting where the border line intersects the scale; the film of liquid drawn up around the stem by capillarity should be disregarded.
The reading
and not
17.
of the spindle, for example, in Fig. 22, is 20 The scale of the hydrometer is read with
H
15
greater ease when the surface of the liquid is level with the brim of the cylinder. Cylinders of the form designed
by Winter
(Fig. 23) are
convenient for this purpose; any

is
115
17
overflow of liquid displaced by the spindle the circular trough.
caught in
18 10
The same attention must be paid to temperature when the hydrometer is employed as in other methods The Brix spindle is calof determining specific gravity.
ibrated at 17.5
C.,
and unless the solution be
of this
temperature a correction must be applied. A table of temperature corrections for degrees of the Brix scale is
23
given in Table 4 of the Appendix; these corrections are to be added to readings made above 17.5 C. and subtracted from those
made
below.
Brix hydrometers are sometimes fitted with thermometers, a form of which modification is shown in
The advantages of this construction disappear Fig. 24, somewhat when working with turbid liquors, which render the reading of the thermometer difficult or impossible.
solution
For general purposes the temperature of the is best taken by means of an accurately standardized special thermometer.
Volquartz* has constructed a Brix spindle with a correction scale, the mercury of the thermometer in the stem indicating, instead of temperature, the correction
Fig. 24.
Brix
necessary to be added to the scale reading. The method of operation may be seen from Fig. 25. The spindle in
with the illustration indicates 10.0 Brix; the mercury of the thermometer, thermometer marks 2.7; the reading corrected to 17.5 C. 2.7 = 12.7 Brix. If the mercury is below the mark is, then, 10.0
spindle
(17.5 C.), the correction

*
must be subtracted.
47
Vos^tka* has constructed a Brix spindle with movable scale, which after adjustment to the temperature of the sugar solution gives the
true reading directly. For determining the Brix of dilute sugar solutions, an operation of considerable importance in exhausting filter-press cake ("sweetening " " sweet- water off"), a variety of spindles known as ^-^ These hydrometers spindles has been constructed.
have a large body with a thin stem, so that the readThe sweet ings can be easily made to 0.1 degree. filters the has from water as it comes usually a temto prevent the delay perature of 60 to 80 C., and,
incident to cooling the solution to 17.5 C., sweetwater spindles are often calibrated at high temperatures.
80 0.
of such spindle is graduated to read in water at 75 C., and 5 Brix in a degree Brix 5 per cent sugar solution of the same temperature;
One form
such a spindle cannot of course be employed at other

temperatures,
limited.
17 5
so
that
its
usefulness
is
somewhat
Another form of sweet-water spindle (Fig. 26) is Beto 5 Brix in the normal way. graduated from in the continued are the divisions the mark low with scale a double result the same manner, being
the
division in the middle.
jj
At
17.5 C. the read-
ings of the
upper
scale give the true Brix; at temper-
atures above 17.5 C., sweet waters will read less than the true Brix. At 70 C. a 5 per cent sugar solution on the spindle, a 4 per cent solution reads 1, a solution 2 cent a 3 per cent solution 3, a per 2,
Fig. 25.
Volquartz
per cent solution -4, and pure water -5. The true Brix can be determined for any temperature by
1
.
,
spindle with temperature correc-
means
of a correction table; determinations by this instrument can always be controlled by cooling the solution to 17.5 C. Still another form of sweet-water spindle has been devised by thermomLangen. This spindle (Fig. 27) contains within its body a The graduated scale in the stem eter graduated from 30 to 70 C. of Langen's spindle differs from other forms, however, in not giving Brix degrees, but in simply indicating the thermometer reading for each division to which the hydrometer will sink in pure water. If placed, for example, in distilled water of 30 C., the instrument
* Z. Zuckerind.
Bohmen,
27, 689.
48
will sink to the division
SUGAR ANALYSIS
30 on the stem, and in water of 70 C. to thermometer and scale of the give the same readings between 30 and 70 when the
in other words, the
in
distilled
the division 70;

spindle will
instrument
spindle
is
is
floated
water.
When
the
placed in a sweet water, the reading of therscale will no longer agree. The spindle necessarily sinks to a lesser depth than in water, and the scale of the stem gives a dif-
mometer and
GO
ferent reading from that of the thermometer, the difference between the two being proportional to the concentration of
solution.
50
In
40
sweetening off, it is only necessary to observe the readings of thermometer and scale; the
differences
between these decrease as the exis
traction proceeds, until with the coincidence

of the
two readings complete exhaustion
indicated.
Another form of hydrometer which is frequently used in the sugar factory, but to a
much
ized
less
that of Baume.
extent in the sugar laboratory, is This instrument is standardof
salt; the point at obtained by means of distilled water, and the 15-degree mark by means of a 15 per cent salt solution. The
by means
common
is
the top of the stem
interval
between these two divisions
is
then
divided into 15 equal parts, this graduation being extended downwards on the scale as far
as desired.
ments the temperature

specific gravity of
Unfortunately, in the early instruof the water and the
the salt solution were not

re-
correctly obtained, so that the values of the
Baume' scale divisions have been variously

ported by
Fig. 26.
different authorities.
The
so-called
Fig. 27.
Sweet-water
e>
"old" Baume degrees, as calculated by Brix, are stiU used in European countries in the
commercial analysis of molasses * notwithstanding the fact that Gerlach as long ago as
Langen's sweet-water
spindle.
1870 showed the incorrectness of the formulae employed by Brix in his

calculations.
*
"
Friihling's
Anleitung," p. 74.
49
at 17.5 C., 1.11383. mark, in terms of the
Gerlach found as the specific gravity of a 15 per cent salt solution The volume of a Baume" spindle up to the volume of a single scale division, is then equal 1 11383 X 15 The specific gravity S corresponding to any to -T r = 146.78. 1 J..J.IGOO
scale division
of the
'
Baume
It is
scale
can then be calculated by the

of this formula that the so-
formula
called
S =
by use
degrees have been determined. The relationship between percentages of sugar, or degrees Brix, specific gravity and the new and old degrees Baume, is shown in Table 3 in the Appendix.
"
new " Baume
CHAPTER
A SECOND
is
IV
method
by means
of the refractive index.
of estimating the percentage of sugars in solution The general applicability of this

specific gravity,
method, as in the case of

solutions of
all
sugars of equal concentration index of refraction.
depends upon the fact that have nearly the same
If a beam of light from one medium, such as an inclined angle upon the surface of a second medium, such as water, it will be found that the beam upon entering the second medium is bent or deflected from. its original course. A good example of this phenomenon, which is called refraction, is the bent appearance There is a of the oar of a boat when seen obliquely under water. general law of refraction for all transparent liquids and solids which may be stated as follows: For two given media and the same ray of
Law
of Refraction.
air, fall
at
light
(same wave length), the ratio of the sine of the angle of incidence to the sine of the angle of refraction is always a constant quantity for
the same temperature. In Fig. 28 and m' are two media; PP f is drawn perpendicular to the dividing surface FF'. in the Let a beam of light pass through direction LO; a part of the beam at the point of the surface is re-
flected in the direction
OL'; another part of the
beam
entering m'
falling
is
refracted in the direction OS.
The
angle
LOP
which the
ray
makes with the perpendicular is the angle of incidence, or i; the angle SOP' which the refracted ray makes with the perpendicular is the
angle of refraction, or
refraction.
r.
The
ratio
smr
is
called the index of
This ratio in Fig. 28

sin i
-
is
represented by
-r-
line cd
The
media.
ratio If v is
smr
If
is
also that of the velocities of light in the
two
then
the velocity of light in the refracted ray
m and
v'
the velocity in
f
,
S1T1
W
-
sin
is
bent toward the perpendicular
as in Fig. 28, the velocity v' is smaller than v, and the medium m' is called of greater optical density than m. Optical density must not be
50
51
nnnfiiser confused
all
with material density, since the two expressions do not at
correspond. If the ray of light in Fig. 28 pass from a denser medium m! into a in the direction SO, it will be refracted in in the rarer medium
direction
OL.
In this case the index of refraction
is
sin
.)
which
is
the
reciprocal of the index for light passing in the opposite direction. The refractive index varies with the wave length of the light, increasing
Fig. 28.
Illustrating
law of refraction.
from the red towards the

follows that
violet
when ordinary light of the different prismatic colors; this unequal refraction for light of different wave lengths is called dispersion.
Measurement
solution can be
of Refractive Index.
end of the spectrum. From this it is refracted it is decomposed into light
The
refractive index of a
measured in a variety of ways. One of the simplest methods, which is of more value for demonstration than for accuracy, This apparatus, shown in is by means of the refractometer trough.
inner curved surface of Fig. 29, consists of a semicircular trough, the the of side The which is divided into degrees. trough corresponding
to the diameter of the circle consists of a plate of glass which is made a narrow perpendicular slit at the center c.
nontransparent, excepting If the trough be filled partly with a solution and a
beam
of light fall
slit
upon the glass, that part of the
beam
passing through the
above
52
SUGAR ANALYSIS
the surface of the liquid will mark the angle of incidence and that part passing below the surface will mark the angle of refraction. In the
Fig. 29.
Measuring refractive index by refractometer trough.
illustration,
where water
is
used, these angles are 60 degrees
and 40
degrees respectively.
sin sin
60 40
0.8660
0.6428
1.34 or n, the approximate index of refraction.
Fig. 30.
Illustrating principle of total reflection.
In the construction of refractometers for more accurate measurements, instrument makers generally employ the method of total reflection.
The
principle of this
Let
m and mi
method can be understood from Fig. 30. be two media, such as glass and water, of which m is
53
the more optically dense, the dividing surface being SF. The beams of light which fall from the source L upon SF at various angles are
refracted, in
mi in different directions.
in the
fracted
and proceeds
same
direction;
The beam LO the beam

ot,
J_
SF
is
not re-
Lo,
making the
angle of incidence i, refraction r; in the
is
refracted in the direction

is
same way Loi
refracted to
making the angle of Oiti, and Loz to O-&L.
As the angle of incidence for the falling beam increases, there finally comes a point at o 3 where the refracted ray o 3 Z 3 coincides with the surIf the angle of face SF, and the angle of refraction r 3 = 90 degrees. incidence be increased beyond i 3 to it, the beam which previously was only partly reflected is totally reflected in the direction 2 4 and there is
,
no refraction in m\.
a
.i
Since
1*2
>
smr 3
the index for the
beam before
total
sin
reflection, equals-;
sin r2
etc.,
sini
-:
sin r
n,
and
since sin r 3
i
= ô 90 = 1,
=
n.
it is
evident that for the border line of total reflection sin
In other
words, the sine of the angle of incidence for the border line of total reIt is seen from the diagram that flection is equal to the refractive index.
total reflection
can only take place when
light passes into
an optically
is
is
rarer
medium.
For absolute measurements the refractive index of a substance referred to a vacuum. Since, however, the absolute index of air
only 1.000294, refractive indices referred to air are sufficiently exact In the case of three media such as air, glass, and a for most purposes.
if the index from air to glass be g i, ag and from glass to liquid The sine of the then the index from air to liquid a g i. ag X angle of incidence for the border line of total reflection between glass and a given liquid, multiplied by the index of refraction between air and glass, will give the index of refraction for the liquid with reference
liquid,
to
air.
ABBE REFRACTOMETER
best general instrument for determining the refractive index of The essential part of the solutions is that of Abbe (Fig. 31). sugar of refracand Abbe refractometer consists of two flint-glass prisms
The
To open 1.75, each cemented into a metal mounting. the prisms the latter are rotated on their bearings to a horizontal position with the prism B uppermost; the clamp v is then released and prism B swung on its hinge C. A few drops of the solution to be extive index
nD
open amined are then placed upon the polished inner surface of the fixed is prism A next to the telescope, and prism B, whose inner surface
54
SUGAR ANALYSIS
Fig. 31.
Abbe
refractometer.
55
ground, brought slowly back and clamped as before. The instrument is then swung into an upright position and light reflected from the mirror R upon the surface of the lower prism.
In the following diagram (Fig. 32) FDE and ABC are longitudinal sections of the two prisms in an Abbe refractometer between whose
hypotenuse surfaces
FE
and
AB
(separated by about 1.5
mm.)
is
the
P'
Fig. 32.
Illustrating principle of
Abbe
refractometer.
of light passing from are resolution the of through the lower prism to the surface of the index refractive the to fracted or totally reflected, according film of liquid to be examined.
The beams
AB
liquid.
the hypotdiagram the beams which fall upon re10 line the undergo than inclination enuse surface AB at a less sets the the fraction in the liquid, and, passing through upper prism, of parallel rays s, s', s", etc., are condensed by ,u,u', u",
As shown
in the
the objective
of the telescope upon the
field
XY.
The beams
in the
56
prism parallel to
SUGAR ANALYSIS
10
are refracted along the surface
BA
of greater inclination totally reflected; since these beams the surface of the upper prism, a part of the field
and the beams do not reach

remains in
XY
shadow.
The
sector
telescope of the refractometer (F in Fig. 31)
is
attached to a
S and the prisms to a movable arm J (the alidade) which carries a magnifying lens L. By moving the alidade until the intersection of the
reticule in the telescope field (Fig. 32) cuts the dividing line between the bright and dark portions of the field, the refractive index can be
read directly upon the scale of the sector by means of the lens. The relation between the angles of incidence and refraction of light between air and prism, and prism and liquid, in the Abbe refractometer
may be understood from Fig.
BC and DE of
planes
32. Let PP' be drawn J_ to the end planes f the double prism, and hh be drawn J_ to the hypotenuse
AB
b
and EF.
angle of incidence from air and
angle of refraction in glass; then
for prism,
Let a
= =
sm
=n
which
for the flint glass of the
Abbe mstru-
ment
Let r = a! = b =
r
is
about
1.75.
angle of prism.
angle of incidence in glass upon surface

angle of refraction in liquid
total reflection.
AB
and
line of
90 degrees for border
In
A BOIZ. r + Z BOI + Z BIO =

Z BOI + Z a'
+ Z BIO + Z b = whence r =
2rt. Z's;
rt.
Z's;
a'
+ b.
and
r,
By way
of illustration the following values are given for a, b,
with water as the liquid between the prisms: a = 18 32'.

b
r
= =
10 28'.
60
00'.
E& = al8l7 = L75 = * for

a'
sin
0.3179
air to
pnsm
'
=
=
60
=
10 28'
49
32'.
sin a'
T7
0.76
= 0.76 = n
=n
for glass of prism to water.
1.75
0.76
1.33
for air to water.
57
Each division, therefore, upon the sector of the refractometer representing refractive index is equal to the sine of the angle of incidence in the prism for the border line of total reflection multiplied by the refractive index of the prism.
Since total reflection can take place only
when
light passes
from an optically denser to a rarer medium, the
capacity of the refractometer is necessarily limited to solutions of smaller refractive index than 1.75.
A
light
pensator.
second important feature of the Abbe refractometer is the comThe function of this is to correct the dispersion which white
undergoes in the double prism. Without the compensator the border line between the light and dark parts of the field, owing to the
unequal refraction of light of different wave lengths, assumes the appearance of a band of prismatic colors, which it is impossible to use for
purposes of adjustment.
The compensator
of the telescope
of the refractometer
is
placed in the prolongation
tube between the objective and the double prism. It consists of two similar Amici prisms, such as are used in a direct-vision spectroscope, and which give no divergence for the yellow D line of the spectrum (i.e., the emergent D rays are parallel with the optical axis).
The two prisms are rotated simultaneously in opposite directions by means of the screw head (Fig. 31). Trapezoidal sections of the two Amici prisms are shown in Fig. 33. Each prism consists of a combination of two crown-glass prisms, with
Fig. 33.
Illustrating principles of compensator.
a third right-angled flint-glass prism between them in the manner shown. If a beam of white light LT fall upon the surface of the first
decomposed into its colored constituents, as shown by the divergent broken lines. In their passage through the prism the red at y, rays are refracted least and emerge at r, the yellow rays emerge the If v. at light refracted are and the violet rays, which most, emerge A'B'D'E' second a enter now ABDE prism from the emerging prism A and A' being to the first (their refracting edges
prism AB,
it is
similarly placed
parallel
prism
and on the same side of the optical axis LL'), the colored rays will emerge from the second prism at the points r', y', and v' respectively, the angle of dispersion for any two differently colored rays being twice
that for the single prism
ABDE.
58
SUGAR ANALYSIS
If the two Amici prisms be now rotated in opposite directions around the optical axis LL', the dispersion of the compensator will diminish until, when each prism has rotated 90 degrees (the difference
from the previous position being 180 degrees), the dispersions of the two prisms neutralize one another and the dispersion of the compenIn this position the refracting edges A and A of the sator is zero. two prisms will again be parallel, but on opposite sides of the optical
f
If we now imagine the direction of the colored rays through axis LL'. the two prisms to be reversed, we have an exact representation of the work performed in the compensator. The band of colored light from
the double prism of the refractometer, passing in the direction L'L, emerges at T as a colorless beam, and the bright and dark halves of the
field are
sharply divided.
sator can be given
examined for Amici prism.
By rotating the screw head the compenan equal but opposite dispersion to that of the liquid any value from zero up to twice the dispersion of a single
After setting the compensator to the point where the colored bands disappear, the reading of the scale upon its drum (T, Fig. 31) enables one to calculate the dispersion of the liquid examined for the F and C
rays of the spectrum, the tive index for the F and
mean
nc (difference in refracdetermined with the help of a rays) being

dispersion np
defi-
special table supplied with the instrument.
Duplicate readings upon the Abbe refractometer with a sharp
nition of the border line should agree within two places of the fourth decimal. After each determination the prisms should be cleaned with
paper and then wiped dry with a piece of soft linen. Abbe Refractometer. For illuminating the refractometer ordinary daylight may be used, in which case the instrument should not be placed in the direct light of the sun. Since, however, daylight (especially in winter) is of variable intensity, and upon dark days not strong enough for the examination of deep-colored solutions,
wet
filter
Illumination of
it is
better on the whole to use artificial light of constant intensity.
An
incandescent electric lamp or Welsbach gas burner is a most convenient method of illumination. A large sheet of cardboard, placed in front of the instrument so as to shield the light from the upper prism
and from the eye
of the observer, will protect the field of vision from the disturbing influences of extraneous light and increase to a marked extent the sensibility of adjustment.
Regulation of Temperature in Abbe Refractometer.

the temperature, the index decreasing as the temperature
The
rises.
re-
fractive index of sugar solutions, as of all other substances, varies with

It is

therefore important in
59
all refractometer work that the temperature be kept constant during the course of observation. In the Abbe refractometer shown in Fig. 31 water of constant temperature is allowed to circulate in the direction of the arrow through the metal casings which
surround the prisms;
a thermometer screwed into the upper casing
indicates the temperature.
Zeiss Spiral Heater and Water-pressure Regulator. A convenapparatus for controlling the temperature of refractometers is the Zeiss spiral heater and water-pressure regulator. This
ient piece of
apparatus shown in Fig. 34 consists of a constant-level reservoir A connected by rubber tubing to the water supply and attached to a The water sliding frame which can be adjusted to different heights.
passes from the reservoir to the spiral heater, which is placed upon a The heater consists of about 12 feet level below the refractometer.
of copper tubing
is
wound
in a spiral
and inclosed
in
a metal jacket which
heated by a Bunsen burner. The water flows from the heater upward to the prisms of the refractometer and thence to a constant-level vessel
The water, which B, from which the overflow escapes to a drain. should not flow too slowly, is first warmed to the approximate temperature by regulating the flame of the burner; the exact adjustment
is
then
made by varying
or lowering the pressure reservoir

of course that
the speed of the flow, which is done by raising on its sliding frame. In this manner
the temperature can be maintained for hours within 0.1 C., provided no variations take place in the temperature of the main
water supply. Instead of the Zeiss heater a large insulated heatable tank holding 50 to 100 liters of water may be used. The adTesting the Adjustment of the Abbe Refractometer.
justment of the Abbe refractometer can be tested by means of liquids or glass test plates of known refractive power. Freshly distilled water = for free from air (n testing the lower divi1.33298) is convenient
sions of the sector scale;
venient for
monobromonaphthalene (n= 1.658) is contesting the upper part of the scale; the latter substance
unless freshly prepared usually requires to be redistilled (boiling point 277 C.). The Abbe instrument is supplied with a glass test plate
whose index
is
of using the plate,
marked upon the upper ground surface. The method which can be applied to any transparent solid, is
that of grazing incidence (explained in detail under the immersion

refractometer).
In using the test plate the instrument is reversed as shown in Fig. 35, the double prism spread open, and the polished surface of the plate
60
SUGAR ANALYSIS
[51
Befractometer X, Prisms
Fig. 34.
Zeiss spiral water-heater with pressure regulator.
61
attached to the upper prism by the capillary action of a drop of monobromonaphthalene; the polished end surface of the test plate is directed
downwards
to receive the reflected rays from the bright inner surface
of the metal casing surrounding
the lower prism.

of several readings
The average
is
taken, the clean and the being wiped prism after reattached each measplate
urement.
Care must be exer-
cised not to confuse the reading in the reversed position of the
The average of the should not differ more readings than two points in the fourth
sector scale.
upon the
decimal from the value marked Should greater plate.
differences
than this occur, the
refractometer should be adj usted.
In some of the instruments the
adjustment
,
is
.
made by moving &

,,
,
the index of the
Fig. 35.
sector
scale
Verifying adjustment of refractometer by test plate,
with a setpin until it corresponds to the value marked upon the test plate. The border line of the field must remain meanwhile upon the intersection of the reticule, so that care must be exercised not to ^disturb the alidade while making
the adjustment. In more recent forms of the
Abbe
made by moving
the reticule instead of the index.
refractometer the adjustment is The process is the
reverse of that previously described. The alidade is first moved until the index of the scale corresponds to the reading of the test plate; then by means of a key the screw (Fig. 31), which moves the reticule, is
turned until the intersection of the cross threads coincides with the border line.
REFRACTOMETER TABLES FOR SUGAR SOLUTIONS
A number of tables have been constructed which give the refractive The first of indices of sugar solutions for different concentrations. such tables was published in 1883 by Strohmer,* who showed also that
a fixed relation existed between the refractive index and specific gravity
Oest. Ung. Z. Zuckerind., 12, 925; 13, 185.
62
SUGAR ANALYSIS
method
of sugar solutions. Using the 5 culated this relation to be n

specific gravity of
1.00698
of least squares, 0.32717 d, in
Strohmer calwhich d is the
the solution at 17.5 C.

reflactometer,
In 1901 Stolle,* using a Pulfrich

for sucrose, glucose, fructose,
constructed tables
that but very

is
little
a comparison of which showed variation existed in the refractive index of solutions
and
lactose,
of different sugars for the
made up from
same concentration. The following table the observations of Stolle upon sucrose solutions of
different concentrations.
TABLE XIV
Giving Index of Refraction of Sugar Solutions
Concentration,

TABLE
63
XV
C. to constant weight.)
Giving Index of Refraction of Various Sugar Solutions of Different Concentration
(Dried in
Index of refraction, 20 C.
vacuum
at 70
64
SUGAR ANALYSIS
The
table of
refinery products except the very lowest.
Main
(Table
5,
Appendix), which agrees almost exactly with that of Tolman and Smith, The indices give is the one employed by most sugar chemists at present. the percentage of water to 0.1 per cent from 100 per cent to 15 per cent; the percentage of water subtracted from 100 gives the corresponding * percentage of total solids. Stanek has prepared a table of temperature corrections for the table of Main, the figures of which show, as
was indicated by Tolman and Smith, that the temperature corrections for specific gravity and refractive index are virtually the same (Table 6,
Appendix) Schonrock f of the Physikalisch-Technische Reichsanstalt in Berlin has made the most recent measurements of the refractive indices of sugar solutions. A preliminary report of Schonrock's determinations,
.
which as regards attention to scientific detail are probably the most carefully conducted of any measurements thus far made, is given in Table XVI, in which n is the refractive index at 20 C. for the two D lines of sodium light (589.3 ///*) and w the water content of the solution.
TABLE XVI
Giving Refractive Index and Water Content of Sugar Solutions
<
65
The use of the Abbe refractometer was extended to raw sugar cane products by Prinsen Geerligs and van West* who made a special study of the effect of impurities upon the refractive index of sugar solutions. Their results, in connection with observations upon low-grade Java molasses, show that the refractive index of impure sugar solutions is a much truer measure of the actual amount of dry substance present than the specific gravity. The refractometer table (Table 7, Appendix) of
Geerligs f
is
established at 28 C.
and
is
the one best adapted for tropi-
temperature corrections which accompany the table have a range from 20 to 35 C. When corrected to 20 C., Geerligs's results are identical with those of Tolman and Smith, and Main.
cal countries; the
The use
of the refractometer in the
examination of sugar-beet
products has been studied by Lippmann, Htibener, Lange, and many As in the case of sugar-cane products, the refractometer gives others. values for solid matter much closer to the true dry substance than
specific gravity.
The percentage
have partly
be with a
made known amount

Example.
dry matter in sugar products which as such massecuites, moist sugars, etc., can crystallized, the refractometer after dissolving all soluble matter upon
of moisture or
of water.
c.c.
10 gms. of massecuite were dissolved in 10
of hot distilled
water, the weight of mixture after cooling to 20 C. being brought to 20 gms. by addition of distilled water of 20 C. The refractive index of the mixture
was
1.4107, which according to Main's table indicates 54.45 per cent water. 10 (gms. 10.89 54.45 per cent of 20 gms. = 10.89 gms. water in mixture. water added) = 0.89 gm. water in original massecuite, or 8.90 per cent.
Hardin has made comparative determinations of the moisture in different grades of sugar by drying and by the refractometer with the
following results:
Grade
of sugar.
66
SUGAR ANALYSIS
The
variations in the results
tions,
and may
by the two methods are in both direceither been due to the presence of trash in the have
sugar or to the influence of non-sugars. Since the refractometer only indicates the percentage of dissolved solids, any insoluble matter which is present in the weighed sample will introduce an error in the
calculation.
Insoluble suspended matter in sugar solutions, if present in large amounts, will darken the field of the refractometer and interfere with
the adjustment of the border line.
In such cases the solution must
be
filtered.
fractometer.
Examination of Dark-colored Sugar Solutions with the ReIn the examination of dark-colored sugar solutions,
molasses, sirups, extracts, etc., by means of the refractometer, it is not always possible for the compensator to eliminate completely the effects
of dispersion; the border line of the field is then more or less blurred and a sharp adjustment to the intersection of the reticule becomes a matter of some difficulty. In solutions which are not too strongly
colored this trouble may be remedied by bringing the border line to the point of intersection alternately from each side of the field; the average of the readings thus obtained will correct to a large extent the
Some authorities have recommended of faulty adjustment. with dark solutions to adjust the compensator to a colored border, selecting the color most sensitive to the observer's eye; this, however,
errors
is
not very satisfactory, and
if
the blurring of the border line
is
the color of the solution

or clarification.
must be reduced by some method
excessive, of dilution
In the dilution of impure sugar products with water an error will be introduced in the refractometer reading in the same manner as in the determination of specific gravity, owing to the difference in contraction
between solutions
of sugar
and
of the
accompanying impurities
(page 35).
study of the errors resulting from unequal contraction, when

is
dilution
analysis, has
employed in densimetric and refractometric methods of been made by Stanek.* Fifty per cent solutions of betaine and of various organic salts of sodium and potassium were prepared. These solutions were then diluted with known weights of water and the per cent of dry substance determined from the degrees Brix, from the refractive indices according to Main's table, and by drying on sand in a Soxhlet oven at 102 C. A few of the results are given in the
following table:
*
Z. Zuckerind.
Bohmen,
34, 5.
PRINCIPLE
AND USES OF THE REFRACTOMETER

TABLE XVII
67
Comparative Determinations of Solids by Brix, Refractometer and Drying at 102
68
SUGAR ANALYSIS
being higher according to the amount of water added. The usual effect of this contraction is to make the error in estimating non-sugars Neither of less by the refractometer and greater by degrees Brix.
these methods for estimating non-sugars approaches in point of accuracy the method of actual drying.
index of dark impure from dilution with water, may be largely sugar solutions, resulting eliminated by employing the method of Tischtschenko,* which consists in reducing the color of the product by means of a solution of pure The sucrose of about the same density as the liquid to be examined.
errors in determining the refractive
The
in this
disturbing influences of color dispersion in the refractometer field are way overcome without the errors of contraction. The method
is
of operation
etc., is
as follows:
A known
weight
(a) of
the molasses, sirup,
intimately mixed with a known weight
whose sugar content (p) has been the refractometer. The refractive index of the mixed solution
(6) of pure sugar solution, previously determined by means of
is then determined and the corresponding percentage (P) of dry substance found from the table. The percentage of dry substance (x) in the molasses, 6)P, sirup, etc., is then calculated by the formula ax bp = (a
whence
Example.
(a -
+ b)P-bp a
(a)
(6)
Weight of beet molasses Weight of sugar sirup

Sugar in sirup ngof mixture
Solids of mixture
(p)
= = =
14.1028 gms. 13.2438 gms.

51.3 per cent. 1.4538 = 34.87 per cent water
=
(P)
(Main's table).
= 100-34.87 =
65.13 per cent.
78.12 per cent solids in molasses. Substituting these values in the formula, x The method by water dilution gave 79.11 per cent. Direct determination by
drying gave 77.80 per cent. If a sugar sirup of greater density had been used for mixing, the value of x would have been more close to the result by direct determination.
If equal weights of molasses and sugar solution are used in Tischtschenko's method, then a = b .in the formula, whence x = 2P p; the labor of calculation is thus considerably reduced. In using the
method, the mixture of molasses and sugar solution must be perfectly homogeneous. Care must also be exercised, as in all cases, that no air bubbles are inclosed with the liquid between the A comprisms.
*
69
parison of results in determining dry substance in different samples of beet molasses by various methods is given by Lippmann* in the following table:
TABLE XVIII
Comparative Determinations of Solids in Beet Molasses by Drying, Specific Gravity, and Refractometer
Number.
70
SUGAR ANALYSIS
Lead subacetate.

the
71
The principle of the immersion refractometer is the same as that of Abbe instrument, being based upon an observation of the border
In Fig. 37,
line of total reflection.

its
is
immersed in the liquid contained in the refracting surface If V. beaker we suppose light to pass through the top of the glass will prism from the surface A B, the parallel rays sP s'P', s"P",
5
DE
a cylindrical glass prism with
etc.,
Fig. 36.
Zeiss immersion refractometer.
be refracted in the liquid in the direction PM, P'M', P"M", etc. By increasing the angle of incidence for the parallel rays upon the surface DE, a point is reached where the parallel rays rP, r'P' r"P", etc., are This is the borrefracted along the surface of the prism towards D.
,
der line of total reflection as explained under Fig. 30, where the angle of refraction is 90. In the use of the immersion refractometer the
the reversed direction to that just described, being reflected from the mirror through the bottom of the beaker V so as to pass as nearly parallel as possible to the oblique surface of
course of the light
is in
HK
the prism.
The rays
of light
which coincide with the surface
DE form
72
SUGAR ANALYSIS
the border line for total reflection and are refracted upward through the prism as the parallel rays Pr, PV, P"r", etc., which, being condensed
by the
objective
of the refractometer telescope upon the point x of the scale S, form the border line for observation; the rays of
light
which
may strike the prism
M"P"
surface obliquely, as MP, M'P', etc., are refracted in the

,
direction Ps,
PY,
P'Y',
etc.,
and being condensed by the objective between x and y cause

this part of the scale to
minated.
be illuThere being no pos-
sible angle of refraction for light
in the prism greater than that for the border line of total reflection,
the part of the scale

z
between x and shadow.
remains in
refractom-
As
of
in the
Abbe
in
eter, the border line on account
differences
dispersion
is
fringed with color and must be corrected by a compensator in the manner described on p. 57.
The compensator
Fig. 37.
is
placed at
Illustrating principle of sion refractometer.
immer-
(Fig. 38) between the objective and the prism and is ro-
becomes sharp and colorless. scale marks the reading for the whole division; the fractional division is determined by rotating the micrometer screw Z, which controls the scale, until the whole division previously noted is brought into contact with the border line. The reading of the micrometer drum shows the fractional division which remains to be added. Readings can be made by careful observers to agree within 0.1 scale division, which corresponds to 3.7 of the fifth decimal of the refractive index. This exceeds considerably in accuracy the reading of the Abbe refractometer. The adjustment of the Zeiss immersion refractometer scale is made
by the milled ring R until the border line upon the scale The position of the border line upon the
tated
by means
of
distilled water,
which should give a reading of 15 at
PRINCIPLE
17.5
73
C.
The adjustment, however, can be made
at other temperadistilled water:
tures according to the following table. The correctly adjusted refract ometer should
At a temperature
of
show for
74
SUGAR ANALYSIS
adjusting screw inside the micrometer drum and turning anticlockwise, the border line of the field is made to agree with the whole scale division corresponding to the temperature of the water. The loosened micrometer
drum
it
is
now turned
holding
is
firmly in this position, the
retightened.
index marks the proper decimal; nut which governs the micrometer The new adjustment should be controlled by repeated
until
its
readings.
The readings of the Zeiss immersion scale extend from 5 to +105, and are converted into refractive indices or into percentages of sugar
Fig. 39.
Tempering bath
for
immersion refractometer.
by means
Hiibener;
of special conversion tables which accompany the instrument. tables for the immersion refractometer have been prepared by Sugar
these give the sucrose values of the scale from 15 to 106 with percentages of sucrose from 0.00 to 21.71. Each 0.1 division of the scale corresponds to about 0.02 per cent sucrose or other sugar, and readings can be made with this degree of exactness. (See Table 8,
Appendix.)
For controlling the temperature of the water bath, containing the

*
Dent. Zuckerind., 33, 108.
PRINCIPLE
75
beakers of solution for the immersion refractometer, the spiral heater and water-pressure regulator previously described may be used. A
tempering bath holding 10 liters of water and with a revolving frame When the for 12 beakers (shown in Fig. 39) is also recommended. been reached in the has beakers the solutions are proper temperature
read in sequence, the refractometer prism being wiped dry after each immersion. When large numbers of solutions are to be tested, each solution as soon as read is replaced by]a beaker of fresh solution, thus
giving sufficient time for regulation of temperature without interruption
of work.
When only a few cubic centimeters of solution are available or when the liquids to be examined consist of dark-colored sirups, molasses, extracts, etc., the immersion prism is fitted with an auxiliary prism
held in position by means of a metal beaker and cover. The method of use is somewhat similar to that of the Abbe refractometer; the hypotenuse surface of the auxiliary prism is covered with a few drops of
solution
and then inserted
in the beaker against the face of the

is
sion prism so that a thin layer of liquid
immerthe between two. spread
The remarks upon
illumination under the
Abbe refractometer
also
apply to the immersion instrument. As to the particular choice of refractometer for the sugar laboratory,
the chemist
must be guided by
is
his requirements.
The Abbe refractom-
eter has the widest range,
adapted to smaller quantities of solution,
and
more convenient to operate. The immersion refractometer, however, is more accurate in adjustment and much less expensive. For general work the Abbe instrument will be found more useful; for more limited operations upon solutions below 20 Brix, such as beet and
is
cane
juices,
sweet waters,
etc.,
the immersion instrument possesses
certain advantages.
CHAPTER V
POLARIZED LIGHT, THEORY AND DESCRIPTION OF POLARIMETERS
IN order to arrive at a sufficiently clear understanding of the optical which underlie the construction and manipulation of polariscopes, a brief reference must be made to the physical theories of light.
principles
According to the undulatory theory of Huyghens, light consists of wave motions of the luminiferous ether, the imponderable medium which pervades all space and penetrates all matter.
vibrations or
Waves of light, contrary to those of sound, vibrate transversally In Fig. 40 a graphic representation is given instead of longitudinally.
D
Fig. 40.
Illustrating principle of a light wave.
wave vibrating transversally to the direction of motion LM. The plane of vibration of ordinary light takes all possible positions about this line of motion. The distance OB or O D from the middle to the extremity of an oscillation is known as the amplitude of the wave. The distance from A to E (points in the same phase) is known as the wave length (X), which for light is expressed in millionths of a millimeter GU/X). The number of waves per second is called the rate of vibration (N). If the velocity of light through a homogeneous medium
of a light
f
be V, then
N=
Â
According to Maxwell's electromagnetic theory, which has since been confirmed by the work of Hertz, there are two sets of transverse vibrations in the transmission of a ray of light, the one an electric displacement of the ether, and the other a magnetic displacement, the planes of these being perpendicular to each other. The intensity of a ray of light is proportional to the square of the
76

flTYIT
77
amplitude; the color depends upon the rate of vibration of the ether wave. The color of light may, therefore, be expressed mathematically or of its wave length X. The values in terms of the rate of vibration
of
N and X for the average

:
ray in each color of the
spectrum are given
in the following table
TABLE XIX.
Color.
78
SUGAR ANALYSIS
Malus, in 1808, discovered that the polarization noticed by Huygens in Iceland spar could also be produced by reflection. If a beam of light (as LO in Fig. 28) Polarization by Reflection. a surface of fall upon the smooth transparent substance, it is decomposed
and refracted rays. The reflected rays at a definite are completely polarized, the plane of the lines of incidence angle of reflection incidence and being the plane of polarization.* These obserinto reflected
vations, according to Fresnel and Arago, could be explained only by supposing that the vibrations in a light wave are tran verse to the direc-
and that during reflection these vibrations are reduced to a single plane, which is perpendicular to the plane of polarization. The angle of incidence at which reflected light is completely polarized is called the polarizing angle, and varies according to the refractive
tion of motion,
This relationship is expressed by of the reflecting substance. Brewster's law, viz. The tangent of the polarizing angle is equal to the index of refraction for the reflecting substance, or tan i = n. The = 1.54) is accordingly about 57 degrees. polarizing angle of glass (n
power
A simple apparatus for producing The Norrenberg Apparatus. and studying polarized light is that of Norrenberg, shown in Fig. 41. A and B are two mirrors of black glass; the upper mirror B can be rotated by the crank D around the vertical axis of the instrument, the
angular displacement being indicated upon a divided circle S. The planes of the two mirrors are first placed parallel, at an angle of 45 degrees to the vertical, and a beam of light is allowed to fall upon the
mirror
at
an angle
of incidence of 57 degrees.
The
reflected
beam
is
then completely polarized and, passing upward, is reflected from mirror B upon the screen C, where it appears as a bright spot. With the
mirrors parallel, the planes of incidence and reflection, and hence of Without changing its inclinapolarization, coincide for each surface. B mirror with its screen is rotated C the around the by the crank tion,
vertical axis.
The plane
of incidence
and
reflection for the
beams
of
polarized light at mirror
no longer coincide with that at mirror A;
the intensity of the spot of light upon the screen accordingly begins to diminish until, after a revolution of 90 degrees, the screen is perfectly
In dark, all the light being refracted and absorbed in the mirror B. the latter position the planes of incidence, and hence of polarization, for the light of the two mirrors are at right angles, and the mirrors are
said to be crossed.
*
By turning D
in the
same
direction the spot of light
The refracted rays of light are also polarized, but not completely; most of the refracted rays, however, are polarized in one direction, their plane of polarization being perpendicular to that of the reflected rays.
79
mum
reappears upon the screen, and after 180 degrees again reaches maxibrilliancy, in which position the planes of incidence and of polarization again coincide in both mirrors;
at 270 degrees,
is
when
these
planes are again at right angles, the spot of light
reextinguished.
Fig. 41.
Norrenberg's polarizing apparatus.
one of the points of extinguishment of light upon the screen the glass cylinder F containing a solution of sucrose or other optical active sugar be inserted in the path of the light rays reflected from A,
If at
80
SUGAR ANALYSIS
the illumination upon the screen will reappear. The plane of polarization of the light reflected from A must, therefore, have been rotated by the sugar solution through a certain angle in order that reflection could
until the plane of polarization for the again brought perpendicular to the plane of incidence, the point of maximum darkness is reestablished. By measuring upon S the positions of maximum darkness, before and after inserting the
light
take place from B\ by turning
upon B
is
through which the sugar solution has rotated the plane of polarized light can be measured. In the Norrenberg apcylinder, the angle
paratus the mirror A for polarizing the light is called the polarizer and the mirror B for measuring rotation the analyzer.
Polarization
by Double Refraction.
Of the several contrivances
available for producing plane polarized light, a modified crystal of Iceland or calc spar is the only one used in the construction of polariscopes
and saccharimeters.
Calc spar is a clear, transparent mineral which cleaves readily into rhombohedra. If a small object be viewed through such a rhombohedron, the image will be doubled. Rays of light in?
passing through the crystal undergo "double refraction." The phenomenon is noticeable in any position of the calc-spar rhombohedron
except in a direction parallel to the diagonal joining the two opposite
c
Fig. 42.
D
Calc spar rhombohedron.
Fig. 43.
light in calc spar.
>
Illustrating double refraction of
obtuse corners,
optical axis
known
as the optical axis.
Any
plane including the

is
and perpendicular to the face
of the crystal
called
an
is
axial plane or principal section. In the rhombohedron of calc spar, in Fig. 42, the direction
AH
the optical axis. lar to the face

face.
If
The plane
ABHG
(or
any
parallel plane) perpendicu-
AFGD
is
an
fall
axial plane or principal section to that
a beam of light
it is
LA
upon the
surface of such a
rhombohedron
resolved into two rays, the ordinary ray and the (Fig. 43), extraordinary ray ACE. Both of these rays emerging from the crystal are polarized, their planes of polarization being perpendicular to each
other.
ABO

The Nicol Prism.
set of the
is
81
for polariscope construction it
Before a crystal of calc spar can be utilized must be modified so as to eliminate one
component
rays.
The
best
known method
At each
(that of Nicol)
the following: A rhombohedron length is about three times the width.
ABCD
(Fig. 44) is selected
whose
end and
of the crystal,
wedge-shaped sections
BFC
ADE are removed so as to reduce the acute angles DAB and BCD of the axial plane from
71 degrees to 68 degrees. divided by the plane
The
crystal
is
then
FGEH
the two modified end faces.

are then polished
perpendicular to The cut surfaces
and reunited with Canada

the
balsam.*
are
The
sides of the prism thus obtained
afterwards
blackened and
of cork
whole
is
mounted by means
tube.
and wax
in a metal
Fig.
44.
C
Illustrating construction of Nicol
is resolved into two component rays; the component most refracted (the ordinary ray) meets the film of balsam EF at such an angle that it is completely reflected to the side of the prism, where it is absorbed by the dark The other component (the extraordinary ray), whose vibracoating.
Let represent a principal section of the Nicol prism (Fig. 45). A beam of light LT entering parallel to the long sides of the prism
AFCE
prism.
tions are in the plane of the principal section, is less refracted and, passing through the film of balsam, emerges in a polarized condition
Fig. 45.
Illustrating polarization of light
by a Nicol prism.
from the end surface of the Nicol at the point e. With respect to the end surface of the Nicol FCLM (Fig. 44), the electric vibrations of the
emergent light are in the plane of the principal section, i.e., in the direction of the short diagonal FC; the plane of polarization is in the direction of the long diagonal
"Iceland spar
is
LM.
rather friable, and in practice it is found easier to grind away half of the rhomb instead of cutting it, as generally described. The remaining halves " of two rhombs thus Theory of Preston, ground are then cemented together."
Light," third edition, p. 319.
82
SUGAR ANALYSIS
In the discussion of polarized light, it makes no difference which plane is taken for reference, provided it be always the same. In future pages the terms vibrate, vibration, plane of vibration, etc., refer entirely With this to the electric displacements in the transmission of light.
all
understanding, the statement of Fresnel, which is followed in nearly that the plane of vibration of light is works upon polarimetry,
perpendicular to the plane of polarization, confusion.
can be retained without
The Glan Prism.

scientifically perfect
The type of Nicol prism which is the most and the one most used at present in constructing polariscopes and saccharimeters is that
In constructing this prism the opposite obtuse corners of a calc-spar
of Glan.
rhombohedron
c
are cut off
(as
ABCDEF,
Fig. 46)
by planes
PQR
and
STF
perpendicular to the optical axis which From this passes through the point X.
section a rectangular prism
LMNO
is
sawed
out,
which
is
then cut in half
along a plane through
MN.
After pol-
Fie. 46.
Illustrating construction of a Glan prism.
cemented together again by Canada balsam am mounted as in an ordinary Nicol. Th< great advantages of the Glan prism ovei ,. ,, ,, ,. XT the ordmarv Nlco1 are that the ra y s oi
ishing, the cut halves are
.
light enter the prism perpendicular the end surface and at right angles to the optical axis, thus securii the greatest amount of light capacity per unit of length.
PRINCIPLE AND CONSTRUCTION OF POLARIMETERS

Polarizer and Analyzer.
called the polarizer
combination of two Nicol prisms,

constitutes the essential feature of
and analyzer,
every polariscope. The function which these two parts play can b( be understood from the following diagram (Figs. 47 and 48). The polarizer, which is stationary, is represented by the pris
ABCDEFGH,
whose
axial plane lies
through
ACEG.
A beam
of light
entering from L at the point x is doubly refracted; the ordinary rays are eliminated at o, while the extraordinary rays emerge at e, vibratii
in the axial planes of the prism, with the plane of polarization parallel with the plane BDFH. If the emergent polarized light now enter second prism A'B'C'D'E'F'G'H' (the analyzer), which can be rotat
83
about its long axis, its course will remain unimpeded only so long as it can continue to vibrate in the same axial plane. If the analyzer be rotated about its long axis, the light which enters from the polarizer is doubly refracted and only that component which vibrates in the
Crossed
Nicols
Analyzer
Figs. 47 and 48.
Fig.48
Polarizer
Illustrating principle of polarizer
and analyzer.
plane of the principal section emerges. As the analyzer is rotated the intensity of the emergent light diminishes until after a quarter revolution it is completely extinguished; in this position the axial planes
of polarizer
and analyzer are perpendicular to one another and the two
prisms are said to be crossed (Fig. 48). If the rotation of the analyzer be continued, light will again begin to emerge, until after a half-re volution, when
the axial planes are again parallel, the original intensity will be restored. The amount of light
which
will pass
through the
any position of its axial plane with reference to the polarizer may be readily calculated by
analyzer for
referring to Fig. 49.
Let
AB be the axial plane of the polarizer (always

CD
lay off
stationary) and any given position of the axial of the plane analyzer, the two planes forming the
angle
DOB. From
any distance
OP
as the
amplitude of the light
emerging from the
polarizer,
perpendicular to CD; then the line OL represents the amplitude of the light emerging from the analyzer and PL the amplitude of the
erect
light extinguished in the analyzer.
From
PL
extinguished by
analyzer.
As regards the
relation in intensity,
this is proportional to the squares of the amplitudes:
OP
OL
+ PL
84
If
SUGAR ANALYSIS
we
erect
LM perpendicular to AB and
in
call
the intensity of the light
emerging from the polarizer OP, then the intensity of the light emerging and the intensity of the from the analyzer will be represented by
the analyzer by :: OL PL ). and OP are equal when the planes CD and The intensities is zero when the planes CD coincide (parallel prisms) the intensity
light
extinguished
OM
OM MP (OM MP
:
AB
OM
(crossed prisms). construction and principle of the simplest form of polariscope is the can now be understood from the following diagram (Fig. 50). a of A Nicol and is the stationary analyzer conpolarizer consisting
and
AB are perpendicular
The
sisting of a
movable Nicol mounted
in a revolving sleeve;
the angular
Fig. 50.
Showing arrangement
of parts in a simple polariscope.
rotation of
of
A is measured upon a graduated scale S. L is the source monochromatic light which passes through the instrument to the
eye of the observer at E. We will suppose the Nicol A to be crossed with reference to P, the point of light extinction marking the zero point on the scale S. If a tube T filled with a solution of some optically active substance, such as cane sugar, be now placed between P and A,
the plane of polarized light emergent from P will be rotated from its original position and the light will no longer be entirely extinguished
in A. By rotating the analyzer until its axial plane is perpendicular to the vibration plane of the light emergent from T, the point of exThe angular rotation of the solution in T is tinction is again reached.
then determined upon the graduated scale. By continuing the revolution of the analyzer, light will again emerge from the latter, to become reextinguished at a point 180 degrees from the first reading. Owing
to the fact that light rays of different wave lengths are rotated to a different extent by optically active substances (a phenomenon known
it is necessary that the light used in this type of be monochromatic. polariscope Blot's Polariscope. The original polariscope of Biot* (Fig. 51), constructed in 1840, had an adjustable mirror (M) of black glass for
as rotation dispersion),
Ann. chim. phys.
[2],
74, 401 (1840).
85
the polarizer and a modified prism of calc spar for the analyzer (A). The essential features of this early instrument are still retained in
modern polarimeters, although
in a greatly modified form.
Fig. 51.
Biot's polariscope.
Mitscherlich' s Polariscope. Mitscherlich* in 1844 modified the Biot apparatus by discarding the polarizing mirror and arranging the In the Biot polarioptical parts of his instrument as shown in Fig. 50.
scope the end point was marked by total light extinction. But in the Mitscherlich apparatus a vertical black band with shaded margins
marked .the zero

until the vertical
point.
By
band appears exactly
rotating the analyzer gently to and fro in the center of the field, a zero-
point adjustment can be secured with a probable error of
6 minutes.
is
The Biot-Mitscherlich shown in Fig. 52.
polariscope, with position of
its
optical parts,
Sections of the circular scales used upon the Mitscherlich and other polarimeters for measuring the angular rotation of the plane of polarThe scale in Fig. 53 for a ized light are shown in Figs. 53 and 54. small polariscope indicates 0.1 degree and is immovable, the rotation
being indicated by the position of the zero mark of the movable vernier V. In the illustration the zero mark of the vernier lies between the
*
"
Lehrbuch der Chemie" (1844),
1,
361.
80
SUGAR ANALYSIS
of the scale; to obtain the fractions of a 2-degree and 3-degree division zero mark of the vernier and, moving the from degree, one proceeds of the main scale, comes finally to a dividivisions the along
upward
sion which exactly coincides with one of the divisions of the vernier. In
the illustration this vernier division

is
0.4,
on
the-
main
which, added to the reading scale, makes the angular
rotation 2.4 degrees.
For the larger
polariscopes indicating 0.01 degree the
main
scale is movable, the circular rim divided into 0.25 degree rotating
against the fixed vernier, which gives the readings to 0.01 degree. In the
illustration (Fig. 54) the zero of the
vernier
falls
between 13.50 degrees

of
is
and 13.75 degrees; the 0.18 mark

the vernier
division
in coincidence with a
scale.
is
on the main
13.50 -f
0.18
13.68, which
the angular
rotation indicated.
RobiRobiquet's Polariscope. quet increased the sensibility of the

Biot-Mitscherlich polariscope by in-
troducing a Soleil double quartz plate

as the end-point device.
Fig. 52.
The
is
general
in
The
Biot-Mitscherlich
appearance of this instrument, with

position of optical parts,
Fig. 55.
polariscope.
shown
two plates of quartz of equal thickness, one of which rotates the plane of polarized light to the The plates, which are cut perpendicuright and the other to the left.
consists of
lar to the optical axis of the crystal, are
= position of polarizer 6 = position of analyzer c = lever for rotating analyzer I = condensing lens.
Principle of the Soleil Double Quartz

Plate.
The Soleil double quartz plate
cemented together at their edges and carefully ground and polished. If white polarized light pass through such a plate, the rays of different wave length and color will be rotated to a different degree (rotation dispersion), the rays of less
wave length being rotated the most.
For a piece of quartz 1 mm. thick, cut as above described, the rotation will be 15.75 degrees for the red B ray, 21.72 degrees for the yellow ray of sodium, and 32.76 degrees for
87
Fig. 53
Fig.
54
Sections of circular scales of polariscopes.
Fig. 55.
Robiquet's polariscope.
d = e = / = = g h-i = k =
polarizer
condensing lens Soleil double quartz plate

analyzer
telescope lever for rotating analyzer.
88
the blue
SUGAR ANALYSIS
F ray.
For the average ray in the middle of the yellow specis
thickness of the Soleil plate is so chosen that this average yellow ray is extinguished in the analyzer. This corresponds to a rotation of 90 degrees, or to a thickness of 3.75 mm. (90 -5- 24 = 3.75) for the double plate, when the end point is taken
trum the rotation
24 degrees.
The
for parallel Nicols.
If
a plate of the
above description be inserted between two parallel Nicols and examined with
white
will
light,
the color of the two halves

color,
be of a uniform rose
of
the
blending the yellow.

_j_
the spectral colors minus The relationship of the
60
angular rotations for red, yellow, and blue in the two halves of a 3.75 mm.
at the transition point may be seen from Fig. 56. By rotating the the to left the uniform or analyzer right
CYellow-90J
Blue
120
+ 90) Extinguished P late

+120
Fig. 56.
Soleil
Showing principle of
double quartz plate.
rose color of the plate will change, onehalf to blue and the other to red or
>
vice versa.
a solution of an optical active substance be placed in the tube

If
before the analyzer, the equilibrium in color of the transition tint will be destroyed and the two halves of the field will be differently colored.
Rotating the analyzer to the point where the transition tint is again produced will give the angular rotation of the solution. The Robiquet polariscope, which has a sensibility of about db 4 min-
The rotation angle (a) utes, is of course only adapted for white light. of a substance for extinction of the mean yellow ray was expressed
by Biot
The fact that the point j as / (j = French, jaune; yellow). corresponds to no well-defined line of the spectrum makes it a difficult one to verify, and some confusion has resulted from this cause. Landolt
gives for 1
mm.
quartz, a/
24.5 degrees instead of 24 degrees.
The
value
a.j is
sodium).
always greater than aD (the rotation angle for the D ray of 24 5 The relationship given by Landolt is = i ct a D = 1.128 aD
,-
Zi.iZ
'
using the value 24 degrees the factor 1.111.
= l.W5aD
Many
authorities
employ
Soleil
is
In the examination of colored solutions, the transition tint of the double plate is affected to such a degree that a considerable error introduced in the observation. The use of this end-point device is
for the color-blind.
valueless
For these reasons the transition-tint

little
polariscopes are at present but
used.

Jellefs Half-shadow Polariscope.
89
Efforts to obtain a polarization apparatus which would be free from the defects of those previously named led Jellet* in 1860 to the construction of the first half-shadow
In this type of end-point adjustment, which can be polariscope. secured in a variety of ways, the field of vision is divided into two or more parts, which at the zero position of the analyzer have a uniform shade. Rotating the analyzer to the right will cause one section of
the field to
will
lighter; rotation to the left the effect. produce opposite The half-shadow device of Jellet consists of a rhombohedron of calc spar with its ends cut square and bisected lengthwise by a plane
become darker and the other
are then
forming a small angle with the axial plane of the prism; the two halves cemented together in the reversed position, the result being
that the axial planes of each part are no longer parallel but are tilted toward one another at a slight angle. This reunited prism, placed between the polarizer and analyzer with its line of union bisecting the
field,
izer to
field.
causes the planes of vibration of light proceeding from the polarbe slightly inclined towards one another in each half of the
Rotating the analyzer until it is crossed with the polarizer will not produce extinction, but a uniform shadow or penumbra whose depth will depend upon the inclination of the axial planes in the two halves of the Jellet prism.
The Jellet polarizer was modified by an Cornuf by taking ordinary Nicol prism and dividing it lengthwise by a plane passing through the shorter diagonal of the end. A small wedge-shaped section is then removed from each cut surface and the two halves reunited (see Figs. 57 and 58). This "split" or "twin" prism combines the effect of an ordinary Nicol and Jellet prism. The Jellet-Cornu prism was still further simplified by bisecting
Jellet-Cornu
Prism.
The three only one-half of the Nicol prism in the way described. pieces are then cemented together and the prism squared and mounted, with the split half turned toward the analyzer. This form of prism,
sometimes called the Schmidt and Haensch polarizer, was formerly much used in the construction of half-shadow saccharimeters.t The principle of the half-shadow device of Jellet and its modifications may be seen from Fig. 59. Let GO and HO represent the directions of the axial planes in each half of the Jellet prism, forming with each other the angle GOH (the
*
Rep.
Brit. Assoc., 29, 13 (1860).
t Bull. soc.
t
chim.
"
[2],
14, 140 (1870).
Landolt,
Das
optische Drehungsvermogen
"
(1898), p. 307.
90
SUGAR ANALYSIS
half-shadow angle designated by a and made usually not to exceed It will be seen that with the axial plane of the analyzer 10 degrees).
perpendicular to
PO
extinguished in the analyzer;
the light from the polarizer will not be completely a small amount of light will emerge
End
of Nicol prism
before and after

splitting.
Showing construction
of a Jellet-Cornu prism.
BDE and
GK
and
GE and H F,
BDF, wedge
sections removed.
directions of axial plane before cutting. HK, directions of axial planes after uniting cut surfaces.
and field proportional to the amplitudes equality of light in the two divisions of the field constitutes the end point. By rotating the analyzer to the position A'L' perpendicular to HO, the light in the right half of the field will be
from each half of the
(see Fig. 49).
OM
ON
The
Fig. 59.
Illustrating principle of Jellet's half-shadow polariscope.
completely extinguished, and that in the left half will be increased to '; similarly, with A"L" perpendicular to GO the light in the left half of the field is extinguished and that in the right half in-
from
OM
OM
creased from shadow angle
ON to ON'] it is evident from the above that the halfGOH can be measured by the angle A'OA" through which

the analyzer halves of the
Fig. 61.)
is
91
field.
rotated between the points of extinction in the two (For appearance of field at the several points see
There are several types of polariscopes which use the Jellet-Cornu All of these have the advantage that they polarizer for an end point. can be used with either mixed or homogeneous light, but the disadvantage that the half-shadow angle is fixed and cannot be changed to suit the requirements demanded by different kinds of work. The sensibility of the instrument to slight changes of rotation becomes greater
as the half-shadow angle of the polarizer is made smaller; but, on the other hand, the loss of light at the end point produced by decreasing the inclination of the planes in the two halves of the field lessens the
usefulness of the instrument in polarizing dark-colored solutions.
Laurent's
Half-shadow
Apparatus.
To
overcome
the
last-
* in 1877 condefect of the Jellet-Cornu polarizer, Laurent trived an end-point device in which the half-shadow angle could be
named
changed to suit varied requirements. The Laurent polariscope has the ordinary arrangement of Nicol prisms for polarizer and analyzer, the only difference being that the polarizer is attached to a small lever by which it can be rotated through a small angle to the right or left.
The
essential part of the end-point device
is
perfectly plane
and exactly
parallel to its optical axis.
a thin plate of quartz cut This plate,
which must be of specially prepared thickness, is mounted upon glass The rays in such a way that it covers one-half of the field of vision. of light from the polarizer on entering the plate are resolved into two components, one (the ordinary) vibrating in the plane of the optical thereto. axis, and the other (the extraordinary) in a plane perpendicular
rapidly,
The extraordinary component, being less refracted, is transmitted more and the thickness of the quartz plate is so regulated that when
the two components emerge, the extraordinary one is in advance of the ordinary by half a wave length. The thickness of the plate depends
upon the wave length X of the light, which must necessarily be homogeneous. The component rays which emerge from the quartz plate with half a wave length's (or uneven multiple thereof) difference in vibration are resolved by the analyzer into light which at the end point
half intensity as that in the uncovered aband of the field (the loss of light in the quartz plate by reflection inare which The two planes of vibration, sorption being negligible). to reference with and clined towards each other
is
of the
same amplitude and
symmetrically equally the optical axis of the plate, form the angle of the half shadow.
*
The
Dingler's Polytech. Jour., 223, 608 (1877).
92
SUGAR ANALYSIS
from the
be better understood principle of the Laurent plate can
following diagram (Fig. 60). Let bisecting represent the quartz plate with the edge the circular field, being assumed for convenience to coincide with
LMNK
MK
MK
the optical axis of the plate. Let A A' represent the plane of the f analyzer at the end point and PP the plane of the polarizer, the latter with the optical axis MK. Lay off OB as being set at the angle
POM
Fig. 60.
Showing principle
of Laurent's half-wave plate.
the amplitude of the homogeneous light emerging from the polarizer and draw BC A.AA', then OC will represent the amplitude of the
light
(Fig. 49).
is
emergent from the analyzer for the uncovered half of the field The light of amplitude OB upon entering the quartz plate resolved into two components, one of which OF (the ordinary ray)
vibrates in the plane of the optical axis MK, and the other OC (the extraordinary ray) vibrates in the plane OS _L MK. The quartz plate is of such thickness that the extraordinary component entering at the phase
o> is
accelerated in
'.
its
opposite phase
passage one-half wave length and emerges at the The amplitude OC' being equal to OC, the resultant
OB', between OC' and OF, is equal to OB, and the angle B'OM equal to the angle BOM, the two together being the angle of the half-shadow. The light emergent from the analyzer in both halves of the field will
PP' may be
from
therefore be equal in amplitude and intensity for any angle at which set with reference to MK. On rotating the analyzer
its position,
the equilibrium in shade between the two halves will
93
be destroyed (Fig. 61),* the effect being the same as that described under Fig. 59. The Laurent polariscope, which is the standard instrument in France, has the great advantage, over other forms, of adjustable sensibility
being adapted to only monochromatic light.
without change in zero point, but the great disadvantage of It cannot be used with
TI
Fig. 61.
Showing divisions of double

I,
field of
a half -shadow polariscope.
II,
analyzer crossed with left half of field; analyzer crossed with right half of field;
III,
end point.
white light except when adapted to bichromate filtered light for a quartz wedge saccharimeter. With intense illumination and a small
half-shadow angle (the conditions of greatest sensibility for all halfshadow instruments), the average error of observation according to
Landolt
is less
than
minute.
Concentric Half-wave Plate. Pellin has modified the Laurent polariscope by using a half-wave plate of quartz cut in circular or annular
form.
in Figs.
The
field of vision is in this
62 and 63.*
While the concentric
way divided concentrically as shown field may secure a more correct
O
Fig. 62
Fig. 63
field.
Concentric double
Concentric triple
field.
alignment of the eye with the optical axis of the polariscope, it is much more fatiguing to the eye than the ordinary bisected field. The principle of the concentric half-wave plate is the same as that of the
Laurent plate.
In Figs. 61, 62, 63, and 67b the dividing lines of the fields at end point are intensified. With a properly adjusted instrument the dividing lines completely disappear at end point leaving a plain disk of uniform shade.
*
much
94
SUGAR ANALYSIS
In 1880 Lippich* davised a Lippich's Half -shadow Polarimeter. form of polarizer which combines the advantages of adjustable halfshadow and of adaptability to all kinds of light. The Lippich polarizer consists of two Nicol prisms, one large Nicol, which can be rotated about its
long axis according to the needs of sensibility, and one smaller Nicol, known as the " half-prism,"
which
is
is
mounted
in front of the large Nicol so

field.
as to cover one-half of the
The
half-prism
slightly tilted so that its inner vertical edge
forms a sharply dividing line, which can easily be focused by the eyepiece of the instrument
(Fig. 64).
The principle of the Lippich polarizer can be understood by referring to the opposite diagram
(Fig. 65)
:
Let
OP be the plane
of the large Nicol
and
OH
POH
the plane of the half-prism, the included angle being that of the half-shadow a. Let OB =
the amplitude of the light emergent from the large

Nicol. Draw BG _L OH. Then OG will represent the amplitude of the light emergent from the halfIt can readily be seen that with a loss of prism.
Fig. 64.
struction of a Lippich
polarizer
for
Showing con- a part of the light in the half -prism the ampliin the two halves of the field tudeg QQ> and Q
>
double
do not agree when the perpendicular
-.
*.
^ OA
A ,
to the
,,
N = large Nicol;
n = small Nicol or
" half
LM the amplitudes OC and OD are made equal, Z>= margin of diaphragm; m which position the perpendicular OA no longer
prism;
plane of the analyzer bisects the half-shadow a. By rotating the analyzer slightly from L'M' to
F = projection
of field.
bisects a.
The
angle
OA
makes with the bisector
which the perpendicular OA ' will vary accord-
The Lippich polarizer is ing to the size of the half-shadow angle a. therefore not symmetrical, which is a disadvantage, since by changing the half-shadow a to vary the sensibility there is also a change in the zero point of the analyzer. The latter must accordingly be re-
adjusted for each change in sensibility. The relation of intensities in the light emerging from the large and small prisms of the Lippich polarizer is found as follows:
*
Z. Instrument., 2, 167; 14, 326.
95
^=
OG
cos
Z BOG
T/
cos a.
If
/ and /' are the intensities for the large
and small prisms
respectively, then
-j 2
f)C^
o
cos 2
and
I'
I cos 2 a.
OB
(1)
C
Fig. 65.
r~
-M'
Illustrating principle of Lippich polarizer.
The relation between the angle of the half-shadow a and that of the change in zero point 5 may be calculated as follows When the two halves of the field are matched the amplitudes OC = OD and the inten:
sities
OC = OC OB
sin
Z CBO =
sin
Z POA =
sin
gg
OB
= sinOOD =
sin
Z #04
-sin(f
+
).
(2)
s->
(3)
96
Substituting / and I' for
SUGAR ANALYSIS
OB
and
OG we
,
obtain
OD =
sin2
fe
+ A I';
since
OC*
~6T)
for the
matched
field,
we
obtain
(4)
sin2
~
(f
^)
sin 2
(|
cos - sin
+
)
8
sin 2
(|
+
)
cos2 a.
(5)
sin
cos 5
sin = cos 5 cos
+ cos ^ sin 5 cos a.
Dividing by cos ~ cos 8, we obtain
tan =
-
tan 5
=
1
tan - cos a.
2
COS a
+ tan 5 cos a.
tan 3 ^ 2
,
tan
tan ^ -=. 2 1 cos a
a
(6)
In the above calculation only the light extinguished in the small
There are other factors, however, which must be taken into account in the calculation of the true zero-point correction. Schonrock* has shown that 7.5 per cent of the light is lost reflection from the surface of the small Nicol, and that this amount by is increased to 8 per cent or more by the loss through absorption. Equation 1 for intensity would then become
Nicol has been considered.
WO
The value
of 8 thus modified
1
Cl
\J \J
'
would be expressed by
tan
cos
a A/O92
-.
a
(8)
l+cosaVo.92
tan -. 2
Bates f has shown, however, that a part of the light lost by reflection from the sides of the small Nicol is again restored in the analyzer, and
that
when all factors such as depolarization, size, shape, and inclination of the small prism, etc., are taken into account the true value of 8 is
between those calculated by equations 6 and
ing
8, the exact figure dependof each individual Lippich system. Apart from the disadvantage that the zero point must be corrected
upon the construction

*
t Ibid., 68, 821.

for
97
changes in sensibility, the Lippich polarizer
is
the best for general

in rotation.
use and the one
most
sensitive to
minute changes
The
average error of adjustment, according to Landolt, with bright illumination and a half-shadow angle of 1 degree, is only about 15 seconds (0.004 degree).
The sensibilLippich Polarizer with Triple Field. been almost doubled ity of the Lippich polarizer has by using two half-prisms in place of one, the system
being so arranged that the field of vision is divided into three parts (Figs. 66 and 67). ^The principle of the
triple field
can be understood by referring to Fig. 67a.

ac,
Let AC,
Nicol N, and ab
and n' must be perfectly parallel in order that the halfshadow angles a and a' be equal for both half-prisms,
an absolute essential if perfectly uniform illumination It sometimes hapis to be obtained at the end point.
pens that the two half-prisms get out of parallelism through jarring of the instrument or expansion and contraction of the mountings. There will then be Fj 6g Showing g two end points for the half-shadow, according to of construction which side the middle of the field is made to agree. Lippich polarizer
for tri P le fielch then obliged either to take but one end points, which is equivalent to reducing A, large Nicol; the instrument to an imperfect double field, or else to # and C, small half-
and a'c' represent planes of the large and a'b' planes of the half-prisms n It will be seen that ab and a'b' respectively.
The observer
of these
is
readjust the planes of the half-prisms to parallelism, a most delicate as well as time-consuming operation,
in sensibility of the triple field
p^
'^
O f îa
For instruments requiring constant use the increase can hardly be said to
disarrangement. The more point device is much to be preferred for ordinary laboratory conditions.* Lippich Polarizer with Quadruple Field. a polarizer with quadruple field (Fig. 68)
*
offset the increased sensitiveness of
the polarizer to simple double-field end-
phragm; E and F, inner edges of half-prism which form the divisions H, J, and K of the triple
field.
Lummerf has
by
constructed
placing before the larger
chemists wrongly use the expressions half-shade and triple-shade in place of the terms double field and triple field. The term half-shade or half-shadow, (German, Halbschatten; French, penombre), refers to the depth of shade in the field at
Many
the end point and not to the division of the meaningless.

t Z.
field.
The
expression triple shade
is
Instrument., 16, 209.
SUGAR ANALYSIS
n
b
a
Fig. 67.
Illustrating principle of Lippich polarizer for triple field.

I,
II,
analyzer crossed with outer divisions of field; analyzer crossed with inner division of field;
III,
end point.
Nicol
one large half-prism B, and before the latter two smaller halfThe increased complicaprisms C and D. tion of this form of polarizer has prevented
its
general introduction.
Another Wild's Polaristrobometer. form of polarizing apparatus, whose peculiarities of
by by Wild* shown in
rotated
itself, is
construction place it in a class the jpolaristrobometer invented

in
In this instrument, the polarizer (/) is atFig. 69, tached to a divided circle, K, both being
1864.
by a rod and pinion from the screw
the longitudinal axis of the Nicol device placed at e prism. consists of a Savart double plate made up
C around
The end-point
two sections of calc spar each 3 mm. thick, cut at an angle of 45 degrees to the optical axis of the crystal, and cemented
of
together so that their principal sections

cross at right angles. cross threads
is
diaphragm
with
placed in the focus of the
The anobjective lens d of the telescope. a is stationary, being usually alyzer at mounted with its principal section horiFig. 68.
quadfield.
and forming an angle of 45 degrees with the crossed sections of the Savart
zontal
plate
of Lippich polarizer for
ruple
* "
Ueber
em
neues Polaristrobometer," Bern, 1865.

To determine
illuminated at
99
the zero point of the polaristrobometer, which is first with a sodium flame, a tube of water is placed in the instrument and the ocular of the telescope focused sharply upon the cross threads; the field, except near the end point, consists of a series of dark horizontal parallel bands, the so-called interference fringes, which
Fig. 69.
Wild's polaristrobometer.
of the polarizer increase and decrease in intensity; at certain points of rotation the bands gradually become paler until, at the maximum point of brightness, they are suddenly extinguished in
upon rotation
the center of the
field,
edge (see Figs. 70 and and the extinguished part of the field are symmetrically distributed with reference to the cross threads constitutes the end point. In this
leaving only a slightly shaded border at each The point at which the shaded borders 71).
100
SUGAR ANALYSIS
position the plane of the polarizer is parallel with one of the crossed planes of the Savart plate, so that the end point reoccurs every 90 deIn case the extinguished part of the fringes is too wide for grees.
accurate adjustment, the intensity of the light should be diminished until the borders of the fringes are brought sufficiently close to the The fringes haye usually a different appearance at each of reticule.
the end points, and also with colored solutions, so that a beginner must familiarize himself with the various characters of the field before making
Fig. 70
Fig. 71
Showing
field of
Wild's polaristrobometer.
Interference fringes before end point. Fig. 70. Interference fringes at end point. Fig. 71.
In case the zero points of the scale and vernier do not coinreadings. cide at the end point, the deviation may be noted and applied to the readings as a correction, or else they may be set at zero and the instru-
ment brought
into adjustment
is
by gently turning the screw G
until the
proper end point
secured.
If the polarizer is set at one of the four zero points and a tube of sucrose solution be placed in the trough, the interference fringes will
reappear. The polarizer must then be rotated to the left (opposite to the rotation of the sugar solution) until the fringes again disappear. The angular displacement of the polarizer to the left gives the angular
rotation of the sucrose solution to the right.
through a telescope
latter
is
which
is
The readings are made focused upon the fixed vernier J; the
by a flame at Q. The average error of adjustment to Landolt is about 3 minutes. according The divisions of the scale upon the Wild polaristrobometer are made
illuminated
usually in both circular degrees
and
in degrees of a sugar scale giving
percentages of sucrose. The sugar scale is constructed by dividing 53.134 circular degrees into 400 equal parts. Each of these sugar divisions corresponds to the rotation of 1 gm. of sucrose dissolved to
1000
c.c.
and polarized
in a
200-mm. tube; 10 gms.
of pure sucrose dis-

solved to 100
c.c. will
101
indicate the 100-degree point of Wild's scale,
20 gms. sucrose dissolved to 100 c.c. will indicate the 200-degree point, 30 gms. the 300-degree point, and 40 gms. the 400-degree point. The normal weight of the sugar scale of the Wild polaristrobometer can therefore be varied according to the concentration of the product to be examined, the readings obtained with the 20-gm., 30-gm., and 40-gm. normal weights being divided by 2 or 3 or 4, as the case may be. The Wild polaristrobometer, although formerly used in many European laboratories, finds at present but limited application in technical sugar analysis.
DESCRIPTION OF STANDARD
MODERN POLARIMETERS
of a
parts of this chapter will be devoted to descriptions few standard forms of modern polarimeters. As a type of instrument of French manuLaurent's Polarimeter. facture the Laurent polarimeter is shown in Fig. 72.
The concluding
Fig. 72.
Laurent's polarimeter.
A.
A duplex Laurent sodium burner placed 200 mm. from B. B. Illuminating lens. C. Quadrant whose outer circle is divided into circular degrees and whose inner
circle is divided into sugar degrees. D. Diaphragm containing half-wave plate of quartz. E. Light filter consisting of a crystal of potassium bichromate.
102
F.
G.
SUGAR ANALYSIS
Screw for adjustment of zero point. Geared screw for rotating the analyzer and the arm supporting the verniers. The upper vernier on the right is for reading circular degrees and the lower
vernier
upon the
left for
reading sugar degrees.
Bronze trough 600 mm. long for holding observation tubes. M. Mirror for illuminating scale. N. Magnifying glass for reading scale. R. Tube section containing polarizer; the latter can be moved through a small by means angle by the arm K, which is moved by the crank J through the rod If the solution to be examined is but little colored, the lever U of the lever U. With colored solutions U is is raised, which decreases the half-shadow angle. lowered until the half-shadow is increased to the point of greatest sensibility. The zero point should be redetermined after each change in the position of
L.
the polarizer.
The 100-degree point

corresponds to
is
of the sugar scale of the
Laurent polarimeter
an angular rotation of 21.67 degrees (21 40'), which the value given by French authorities to the angular rotation of the
thick plate of quartz cut perpendicular to the optical axis (see page 112). The normal weight of sucrose corresponding to this rotation is given as 16.29 gms. dissolved to 100 c.c. and polarized in the
1
mm.
200-mm. tube.
The sugar
scale extends
400 divisions to the right and
200 divisions to the
left, thus giving ample range for polarizing all If desired, the sugar scale of the dextro- and levo-rotatory sugars. Laurent polarimeter is adjusted according to the so-called Interna-
tional saccharimetric
division of the International
The value of the 100-degree scale of 20 gms. scale in circular degrees would equal
is
21 67
'
20
lo.^y
of the
26.605 degrees; this
trifle
more than the
circular value
Wild 20-gm. scale, viz., 26.567 degrees, the difference being due presumably to the adoption of a slightly different standard value for
the specific rotation of sucrose. Pellin's Polarimeter. Another type of French polariscope is the half-shadow polarimeter-saccharimeter made by Pellin, shown in Fig.
73.
The
prism;
the half-shadow angle
polarizer of this instrument consists of a modified Jellet-Cornu The division of the is therefore fixed.
is
quadrant into circular and sugar degrees Laurent polarimeter.
identical with that of the
The Pellin polarimeter with variable half-shadow angle (Fig. 74) makes use of a half-wave plate of quartz for the end point, which is constructed for either divided or concentric fields. The arrangement of optical parts and method of manipulation are the same as in the
Laurent polarimeter.
103
Fig. 73.
Pellin's polarimeter with Jellet-Cornu prism.
Fig. 74.
Pellin's polarimeter
with half-wave plate.
104
SUGAR ANALYSIS
A simple form of Lippich's polarimeter use is shown in Fig. 75. chemical Angular rotations adapted for general with this instrument to about 0.015 can be measured degree.
Lippich's Polarimeter.
Fig. 75.
Simple form of Lippich's polarimeter.
h.
Lever for moving large Nicol of polarizer and regulating sensibility. The halfshadow angle which is read by the scale can be varied from degrees to 20 decircle for
K. Divided
measuring rotation.
The
circle
with analyzer in
and
The readings of the scale are made telescope at F is rotated by the screw T. on each side of the circle through the lenses I, which are focused upon the fixed
verniers at n.
P. Location of Lippich polarizer. S. Detachable end for holding light
filter.
A form of the Lippich apparatus devised by Landolt for more general

use
is
shown
in Fig. 76.
that any form of tube or container or substance to be polarized.
This instrument presents an advantage in may be used for holding the solution
The trough
of the polariscope for holding ordinary tubes
can be
removed and the support T employed. The latter is raised or lowered by the screw q and moved laterally upon the tracks c. For polarizing
materials in hot or cold condition, the apparatus G, consisting of a
105
Fig. 76.
g.
Landolt's polarimeter for general use.
Lever for rotating circle R', the final adjustment is made by means of the micrometer screw m after fixing the clamp k. P. Position of Lippich polarizer with two half-prisms giving triple field.
Fig. 77.
Large model Landolt polarimeter.
106
polariscope tube in
SUGAR ANALYSIS
an asbestos-jacketed bath, is employed. The plate then removed and the bath placed directly upon the tracks c. The burner for heating the bath is placed upon the adjustable stand under-
T is
The center narrow tube projecting through the replaceable top bath receives the overflow from the observation tube; the other tubes serve for a thermometer and stirrer for the liquid of the bath. For polarizing at low temperature a cooling medium is used in the bath, in which case the ends of the observation tubes must be covered with
neath.
of the
desiccating caps to prevent condensation of moisture

glasses.
upon the cover
A
circle
degree,
type of more elaborate polarimeter, which can be read to 0.01 The divided is the large Landolt instrument shown in Fig; 77. (driven by the wheel T and micrometer screw m) is covered by a
cap K.
this
Small mirrors Si and

is
lamp through openings

instrument
S2 reflect light from the observation in the cap to illuminate the scale. feature of the double trough by which different tubes of solu-
tion can be brought into the field
by movement
of the large lever
H.
VERIFICATION OF SCALE READING OF POLARIMETERS

of the divided circle upon a polarimeter should be by taking check readings at different points upon opposite The division and mounting of the circle in the best sides of the disk. instruments is made with great accuracy, and, unless the disk has been warped or bent, check readings on opposite sides of the circle will agree much closer than the observer can set the scale for a matched field.
verified
The graduations
scale.
Polariscope readings should always be verified upon the opposite It is also well to reverse the circle 180 degrees and repeat the readings each way from the other side. By so doing the observer will
errors
have 4 sets of readings, the mean of which will practically eliminate all due to faulty scale division or eccentricity. The example on page 107 of readings made upon a sugar solution will illustrate the method.
to the point of greatest sensibility, the angle being small for light-colored solutions and Since altering the half-shadow of the Lippich larger for dark liquids.
of the halt-shadow angle
is
The adjustment
made
system produces a change in zero point (p. 95), the adjusting lever should never be disturbed during a set of observations. The analyzer, if desired, can be brought back to the of the scale for any change in the half-shadow angle by means of a small regulating screw (shown at
The better method, however, is to establish the zero point the upon scale, as in the following example, and subtract this from the
a, Fig. 77).
scale reading.
107
CHAPTER
VI

WHILE the instruments described in the previous chapter are adapted to the examination of all optically active substances, saccharimeters are designed solely for polarizing sugars. For convenience
the scale expressing angular rotation is replaced upon the saccharimeter by one graduated according to the decimal system indicating percentages.
THE QUARTZ-WEDGE COMPENSATION

Owing
to the
many
difficulties
and inconveniences connected with
the use of sodium or other monochromatic light in practical work, the French physicist Soleil was led in 1848 to devise a means by which
ordinary daylight or lamplight could be used for measuring the optical rotation of sugar solutions. This invention, known as the quartzfeature of all saccharimeters. is the characteristic wedge compensation,
In the quartz-wedge saccharimeter the polarizer and analyzer are both stationary; the rotation of the sugar solution is measured by shifting a wedge of optically active quartz between the solution and analyzer until the rotation of the wedge system at a certain thickness
exactly neutralizes or compensates the rotation of the sugar solution. By means of a scale attached to the quartz wedge the rotation of the
sugar in solution
is
measured
in percentage.
The
that
i.e.,
compensation is based upon the fact has almost exactly the same rotation dispersion as cane sugar; a section of quartz and a cane-sugar solution of equal rotation for
selection of quartz for
it
have very nearly equal rotations for light Table XX). The small disturbances due to the slight difference in rotation dispersion between sugars and quartz are eliminated by a bichromate light filter. The quartz wedges used in the conSingle-wedge System.
light of
will
one wave length
of all
other
wave lengths
(see
of the quartz crystal;
struction of saccharimeters are cut perpendicularly to the optical axis they may be either of dextrorotatory or levo-
rotatory quartz, the method of mounting the wedge depending upon the character of the rotation. This can be seen more clearly by inspecting the following diagrams (Fig. 78).
108

In diagram
I,
109
R and movable and C wedges The two of different size must have wedges, stationary. wljich though be considered to form dimensions, a may equal angular together single section with sides parallel to the plate A and perpendicular to the The thickness of the two wedge axis of light through the instrument.
is
a fixed plate of levorotatory quartz, and
C two
of dextrorotatory quartz, of
which
is
sections can be increased or diminished

right or
left.
by moving wedge
to the
At the
zero point of the instrument the right rotation of

Levo-rotatory wedge
Dextro-rotatory wedge system
system
II
Fig. 78.
of single
wedge quartz compensation.
the section Imno of the two-wedge system exactly neutralizes the left rotation of the quartz plate A. If a tube of dextrorotatory sugar
placed in the instrument between the polarizer and the compensation plate A, the optical neutrality is destroyed, and it will be necessary to decrease the thickness of the two-wedge section by
sliding
solution be
now
from
o>
towards
'
until the excess of left rotation in
over
If
and C exactly neutralizes the
right rotation of the sugar solution.
the
solution of sugar is left-rotating, it will be necessary to slide B in the opposite direction until the excess of right rotation in B and C over A
rotation of the sugar. In a levorotatory wedge system A is the dextrorotatory and the wedges (diagram II) compensation plate B and C levorotatory, the compensating motion of wedge B being the
equals the
left
reverse of that in diagram I. Owing to the lateral refraction of light from the inclined surfaces of the wedges through the intervening air space (as shown by the
to allow free
the planes of quartz are separated only just sufficiently The circumof the parts without friction. stance that the field is not exactly at the end point, when the thickness
efg),
dotted line
movement
two-wedge section agrees with that of the compensating plate, due to this lateral refraction. The shifting of zero point due to refraction depends upon the wave length of light; the difference in zero
of the
is
110
SUGAR ANALYSIS
point between red light of 760 MM

WJL
wave length, and violet light of 396.8 wave length was found by Schonrock to be 0.059 degree for the
scale.
is
Ventzke sugar
The
scale of the saccharin) eter

is
attached to the large or movable
wedge, and
lating screw.
read by means of a vernier scale attached to a reguIn case the zero marks of the two scales do not agree,
of the field correspond in shade, they can be
when the two halves
brought into coincidence by shifting the vernier slightly to the right or The vernier left by means of a key which fits the regulating screw.
is
never to be
scales are
moved
two
except for making this adjustment, and when the once set has rarely to be disturbed. Owing to the in-
evitable slight fluctuations in the zero point of saccharimeters, it is best to correct the reading by the zero-point error and not to adjust the scale unless there be a persistent difference of the zero point in one
direction greater than 0.1 degree.
The method
of
reading the sac-
charimeter scale can be seen from Figs. 80 and 81. An elaboration of Double-wedge System.
the quartz-wedge
system just described
is
the double- wedge compensation introduced
by Schmidt and Haensch.
The
arrangement of the parts in the double-wedge system is shown in

A
Fig. 79.
c
D
In the double- wedge system the compensation plate is lacking, this being supplied by one or the other of the pair of wedges, which are
The smaller of opposite rotation. are stationary and wedges A and
-n p. Fig. 79.
order to equalize the refraction of the light rays in the air spaces between the inclined surfaces of quartz (as indicated by the
tlon
Showing construction of double wedge quartz compensation.
B and C movaand C are usually mounted with their r points in the same directhe larger wedges
ble.
tem
wedges of each sysEach of the large wedges is as near alike as possible. provided with a scale. These may be read through the same telescope as upon the Schmidt and Haensch saccharimeter (Fig. 80), or by sepaline)
;
dotted
for this reason also the corresponding
are
made
rate telescopes as in the Fric instruments (Fig. 81). In using the double-wedge system for dextrorotatory substances, the scale (Fig. 80) is set at zero with its vernier, and the optical rota-
THEORY AND DESCRIPTION OF SACCHARI METERS

tion
at zero
111
measured upon the scale A; for levorotatory solutions, A is set and the scale employed. An additional advantage of the double-wedge system consists in the fact that any reading obtained upon
PATENT
JOSEF & JAN ERIC
Fig.
80. Scale of double wedge Schmidt and Haensch saccharimeter.
Fig. 81.
K, control scale; A, working scale indicating 85.5 degrees Ventzke.
Scale of Fric saccharimeter with double vernier indicating 97.7 degrees Ventzke (The division betweenscale and vernier is intensified
.
in reality
no dividing
line is seen.)
the working wedge can be immediately verified by removing the tube of solution and moving the control wedge to the point of compensation.
The control wedge under 'such conditions gives the true reading even though the working wedge have a zero-point correction.
Zero-point determination.
Polarization of
directly,
mat
sugar.
Control-wedge
scale.
112
SUGAR ANALYSIS
The zero-point correction of the workbe determined can accurately by taking check readings very wedge ing at different parts of the scale upon the control. By making polarizations in the same way, the local defects of scale or wedge will be almost
Zero-point Determination.
wholly eliminated. The readings in this case are made without removing the tube, the difference between the two scales being the The preceding table, giving the readings upon uncorrected polarization.
the working-wedge scale for various positions of the control, will trate the method.
illus-
THE SUGAR SCALE AND NORMAL WEIGHT OF SACCHARIMETERS

of a saccharimeter scale is usually based upon the rotation of a definite weight (the so-called normal weight) of chemically pure sucrose dissolved in water to 100 c.c. at a specified temperature
The 100-degree point
and polarized at the same temperature
in a
200-mm. tube.
The
greatest
confusion has prevailed in saccharimetry in the past, and unfortunately still prevails, not only as to the size of the normal weight of sugar to be taken for a specified scale, but also as to the conditions of volume and
temperature under which this normal weight is to be polarized. French Sugar Scale. The 100-degree point of the sugar scale of French manufacture is based upon saccharimeters employed upon
the rotation in sodium light of a plate of dextrorotatory quartz 1 in thickness and cut exactly perpendicular to the optical axis.
mm.
The
choice of quartz as a standard proved to be unfortunate, for, owing either to mistakes of polarimetric measurement or to defects in the quartz (through natural imperfection or mistakes in cutting), the
1-mm. plate has been given a different value from time to time, the results ranging from +20.98 degrees, the early figure of Most French authorities at present employ Biot, to +22.67 degrees.
rotation of the
the value +21.67 degrees. The figure, regarded usually as the most exact, is that of Landolt, who, for spectral pure Na light of mean wave
length 589.3 MM, found the value +21.723 degrees. The grams of sucrose necessary to give the same rotation in 100 c.c. as the 1-mm. quartz plate have also necessarily varied; over 20 different values have
been assigned to this quantity, the amounts ranging from 16.000 gms. (Dubrunfaut) to 16.471 gms. (Clerget and Biot). The cause of these great differences is due partly to variations in the quartz standard and partly to variations in the purity of the light used for illumination. The old normal weight established for French instruments was 16.35 gms., and this weight is still largely used in technical work with the Soleil-Duboscq saccharimeter. In 1875 the value of Girard and
113
de Luynes, 16.19 gms., was adopted as the official weight and remained such for more than 20 years, notwithstanding the severest criticism. In 1896 the International Congress of Applied Chemistry at Paris established the value of 16.29 gms. sucrose dissolved at 20 C. in 100 metric c.c., and this is the official weight used at present by the
French Ministry of Finance.
Ventzke or German Sugar Scale. The sugar scale most generally used outside of France and the one employed upon all German saccharimeters is that of Ventzke. This scale as originally devised by
Ventzke
*
1.1 sp. gr.
was based upon the rotation of a solution of pure sucrose of Y^ It was soon found, however, inconvenient, as well as
.
inaccurate, to rimetric work,
make
and the grams
the specific gravity of solution a basis for sacchaof sugar in 100 c.c. of solution 1.1 sp. gr.
was used
metric
for the -normal weight; this
weighed in air with brass weights

c.c.
was determined to be 26.048 gms. and dissolved at 17.5 C. to 100

in 1855
With the introduction Mohr Cubic Centimeter Standard. of the Mohr f cubic centimeter (the volume of 1 gm. of water at
weighed in the
air
17.5 C.
with brass weights), the original normal weight
of 26.048 gms., designed for metric cubic centimeters,
was strangely
enough retained
and used
for determining the 100-degree point of the
sugar scale. In this way the standard was established which up to 1900 was the only one recognized for the Ventzke scale, and which at the present time is still the one most commonly used in commercial
work.
In accordance with this standard, the 100-degree point of the sugar scale is obtained by dissolving 26.048 gms. of chemically pure sucrose (weighed in air with brass weights) in 100 Mohr c.c. at 17.5 C.
and polarizing the same in a 200-mm. tube at 17.5 C. in a saccharimeter whose quartz-wedge compensation has also a temperature of 17.5 C. This normal weight calculated to 100 metric c.c. (volume of
100 gms. water at 4 C.) is equal to 26.048 gms. gms. (1 Mohr c.c. = 1.00234 metric c.c.).
-=-
1.00234
25.9872
Metric Cubic Centimeter Standard.
On
account of the confusion
and mistakes resulting from two standards of volume, the International Sugar Commission, at its third meeting in Paris, 1900, advocated the abandonment of the Mohr for the metric cubic centimeter, and in so
doing also recommended that the temperature of polarization be made 20 C. The change in temperature from 17.5 C. to 20 C. necessitated a recalculation of the normal weight owing to the difference in specific
* J. prakt. Chem., 26, 84 (1842) ; 28, 111 (1843). " (1886), pp. 44-50. f "Chemisch-analytische Titrirmethode
114
SUGAR ANALYSIS
The
is
rotation of cane sugar and quartz at these two temperatures. calculation is made by the following equation, in which 0.000184
coefficient of decrease in specific rotation of sucrose at
the
20
C., 0.000148
the coefficient of increase in rotation due to the effect of temperature upon wedge and scale, and 0.000008 the coefficient for expansion of the
glass observation tube:
2fi
048
1
+(0.000184+0.000148
0.000008)
(20
17.5)}
26.0082
gms. The International Commission decided, however, to make the new normal weight exactly 26 gms., and in accordance with its recommendation the following definition for the 100-degree point of the Ventzke sugar scale has been universally adopted: "The 100-degree point of
the saccharimeter scale
is obtained by polarizing a solution containing 26.000 gms. of pure sucrose (weighed in air with brass weights) in 100 true c.c. at 20 C. in a 200-mm. tube in a saccharimeter whose quartz-
wedge compensation must also have a temperature of 20 C." All saccharimeters using the Ventzke scale are standardized at present in accordance with this definition. According to Bates and Jackson* a solution of chemically pure sucrose under the above conditions gives
a reading of only 99.89 upon the German scale. United States Coast Survey Standard. The old original standard of the Ventzke scale was the one adopted by the Department of Weights
and Measures of the United States Coast and Geodetic Survey, and was employed for many years by the United States Treasury Department in the Custom House laboratories. The 100-degree point of the scale was taken as the polarization of 26.048 gms. (in vacuo) of pure sucrose dissolved to 100 true c.c. of solution at 17.5 C. and polarized at this temperature in a 200-mm. tube. To avoid the labor of reducing
this weight of sugar to vacuo, the flasks
employed
for the
Coast Survey
standard were graduated to contain 100.06 true c.c., the excess of 0.06 c.c. being taken to correct the error of weighing the sugar in air against brass weights. These flasks contain 0.174 c.c. less than the
Mohr cubic centimeter flasks (100.234 true c.c.), which difference, unless compensated, would cause the normal weight of 26.048 of pure sucrose to polarize 0.17 V. too high. To save the operators the trouble
old
of
this correction, the correction of 0.17 was applied to the test The computed quartz plates used for controlling the instruments. values of the Coast Survey test plates were thus 0.17 V. lower than
making
the values marked by the instrument makers for the
Mohr
cubic centi-
meter standard.
*
Scientific Paper,
U.
S.
Bureau
of Standards,
No. 268 (1916).
115
, The policy of the Department of Weights and Measures of the United

States Coast Survey, in adopting a standard different from that in It gave rise to much confusion and miscurrent use, was unfortunate.
understanding, and traces of this confusion still exist, notwithstanding the fact that the United States Bureau of Standards, the Custom House, and all other United States Government laboratories have abandoned
the old Coast Survey standard and now employ the standard of the International Commission of 26 gms. to 100 true c.c. at 20 C. According to the work of both Sawyer* and Rolfe,f who have made
comparative readings of standard quartz plates upon various saccharimeters, there are many instruments in the United States, even of
recent manufacture, which are standardized for a normal weight of 26.048 gms. in 100 true c.c. Whether this condition of affairs is due
to a
mistaken idea of some manufacturers that the old Coast Survey standard is still recognized officially in the United States, is difficult to It is evident, however, that chemists, in order to avoid the consay. siderable errors due to confusion in standards, should state explicitly, in ordering saccharimeters from manufacturers, that their instruments be graduated according to the standard of the International Commission.
When purchasing second-hand saccharimeters, chemists should be particularly careful to subject the same to a thorough examination
and
verification before using.
the
The rotation value of the Ventzke in Circular Degrees. of the scale has been very carefully modern Ventzke 100-degree point determined by Schonrock,t who found it to equal 34.657 circular degrees
Value of
This is the value used at present by pure sodium light. Schmidt and Haensch in the standardization of all their saccharimeters. According to Bates and Jackson (page 114) the rotation value of the normal quartz plate for pure sodium light is 34.620 circular degrees. Bichromate Light Filter. Schonrock|| has shown that in estabof the Ventzke scale by means of sucrose the lishing 100-degree point
for spectral
the white light must be filtered through a 1.5-cm. layer of 6 per cent potassium-bichromate solution in order to eliminate the errors of rotation dispersion between cane sugar and quartz produced by the light
wave length at the violet end of the spectrum. This light has been adopted by the Physikalisch-Technische Reichsanstalt of Germany and also by the United States Bureau of Standards If in
of shorter
filter
*
t t
If
J. Am. Chem. Soc. 26, 990. According to statement in a letter to the author. Z. Ver. Deut. Zuckerind., 64, 521. Technology Quarterly 18, 294. (1905) Z. Ver. Deut. Zuckerind., 64, 521.
||
Upon its certificates for standardization of quartz plates a sugar degree is thus deby the United States Bureau of Standards
"
:
fined
A sugar degree is the one-hundredth
116
SUGAR ANALYSIS
defining the 100-degree point of the saccharimeter scale, and its use is imperative for all accurate work. Many saccharimeters have a 3-cm.
cell,
and
for this length of liquid a 3 per cent
bichromate solution
is
per cent bichromate = 9). For of rotation materials greater dispersion than cane sugar, carbohydrate such as dextrin, commercial glucose, etc., the author has found it
sufficient (centimeter length of cell
necessary to use a solution of double the above concentration (centimeter length of cell X per cent bichromate = 18) in order to secure
constancy of results between different observers for different sources of white light.
In this connection it is important to note that the rotations of the normal weight of sucrose with bichromate-filtered white light and with sodium light, while very closely agreeing, are not absolutely identical owing to the slight differences in optical center of gravity. Measurements by Schonrock* show that, while a normal sugar solution at 20 C. for bichromate filtered white light is exactly equal to the rotation of a quartz plate of 100 V. (34.657 angular degrees), by using sodium light a quartz plate of 100.03 V. (34.667 angular degrees) would
be required.
The
relationship between Ventzke degrees for bichro-
mate
filtered
is
lengths
white light and monochromatic light of different wave seen from the following table :f
TABLE
XX
Showing Rotation of Quartz and Sucrose for Different Kinds of Light
Source of light.
AND DESCRIPTION OF SACCHARI METERS TI THEORY

g It is seen
117
mate
that while the quartz and sugar exactly agree for bichrothe sugar is rotated to a continually greater extent than quartz for light of decreasing wave length. The normal sugar solution, reading 100 V. with filtered white light, was found to read The eyes of some observers 100.12 degrees with unfiltered white light.
filtered light,
are
more
sensitive
than those of others to the disturbances of rotation
unfiltered light is used (owing perhaps to some difference in the pigment of the eye), so that for accuracy and constancy of
dispersion
when
results in all saccharimetric
measurements the bichromate
filter
should
never be omitted.*
Graduation of Saccharimeter Scales.
Manufacturers of sac-
charimeters in establishing the 100-degree point of their sugar scales employ a carefully standardized quartz plate instead of the normal
weight of sucrose.
The
errors
and inconveniences incident
to the
preparation of chemically pure sucrose and to making the solution up to exact volume are thus avoided; the plate, moreover, has the advantage of being a standard which at constant temperature is always unchangeable. Messrs. Schmidt and Haenschf thus describe the method
of graduating the scales of their saccharimeters: " The establishment of the scale divisions of our saccharimeters is
made
After fixing the zero point the linear at a temperature of 20 C. distance of the 100-degree division is determined by means of a normal
quartz plate reading exactly 100 degrees and standardized at the This linear distance is then Physikalisch-Technische Reichsanstalt.
also verified
plates.
divided into 100 exactly equal parts, the intermediary divisions being by means of corresponding normal standardized quartz
surfaces of the quartz wedges are made perfectly plane so that a quartz stratum of half thickness corresponds to a half value in the division. Slight errors cannot be prevented, as it is impossible to
The
obtain quartz wedges of the necessary length which are absolutely The variableness in the specific optically homogeneous throughout. concentration of solution is not taken into conrotation of sucrose with
sideration in the establishment of the scale division, and this must be corrected for by calculation. Aberrations in the scale division caused by
impurities in the quartz can be detected by the control observation tube." The view that the Ventzke scale of modern saccharimeters is cor-
rected for variations in specific rotation of sucrose with concentration, * At its New York Meeting (Sept. 10, 1912) the International Commission adopted the following resolution: " Wherever white light is used in polarimetric determinations, the same must be filtered through a solution of potassium bichromate of such a concentration that the percentage content of the solution multiplied by the length
of the
t
column of the solution in centimeters In a letter to the author.
is
equal to nine."
118
either
SUGAR ANALYSIS
by curving the surface
of the quartz
wedges or by unequal spacby the above statement. A table has been calScale Concentration Reading. upon Effect of culated by Schmitz* to correct for the changes in specific rotation of sucrose through varying concentration, which gives the actual sucrose value of each scale division of the saccharimeter. These corrections,
ing of the scale divisions, is not substantiated
0.0084153 c, which were calculated by Schmitz's formula, [a] D = 66.514 would seem in light of more recent work to require considerable modiThe formula of Landolt, fication.
[afS
66.435
+ 0.00870 c - 0.000235 c
2
,
(c
to 65),
calculated from the combined observations of Tollens, and of Nasini and Villavecchia, is regarded as the most accurate at present (see page
In the following table the author has recalculated the sucrose 176). values of the Ventzke scale for different concentrations, using Landolt's formula. The values of Schmitz are also given for comparison.
TABLE
XXI
Showing Effect of Concentration of Sucrose upon Saccharimeter Readings
Scale division.

wi It will
of the actual sucrose value
119
be seen from the preceding table that the greatest deviation
from
its scale division

is
equation
is
is
only 0.01 V., which
according to Landolt's too small to be detected by the

error according to Schmitz
ordinary saccharimeter. 0.08 V.
The maximum
riinetric
As regards the concentration of sucrose employed in ordinary sacchawork, the variations due to changes in specific rotation may
therefore be safely disregarded. which are distributed both above
The
small extent of these variations,

scale division, justifies
and below the
the policy of the manufacturers in neglecting this factor lishing the divisions of the saccharimetric scale.
when
estab-
VERIFICATION OF SCALES OF SACCHARIMETERS
IImounted
account of the optical imperfections which quartz wedges occait is important that every user of a saccharimeter should verify the accuracy of his instrument. Owing to the fact that the quartz parts of the saccharimeter are
sionally possess,
On
close to the objective of the telescope, the very local imper-
telescope
wedge system are fortunately unnoticed, since, when the focused upon the polarizer, the cone of light rays emanating from the different parts of the field covers an area of the compensator
fections of the
is
equal to the aperture of the analyzer diaphragm (about 6 mm. diameter) and thus distributes and neutralizes any slight local errors due to defects
Such defects in the fixed part of the system (small wedge and compensation plate) are of no account, since the rotatory power of this remains constant; the predominant optical defects of the large movable wedge are the only ones which vitiate the results of observation. Since local optical impurities in the large wedge are diffused over a
of the quartz.
considerable area, for the reason given above, the errors in the saccharimeter scale never consist of sudden jumps, but only of gradual
undulations. It is unnecessary, therefore, as Landolt has shown, to standardize every division of the scale. The errors at every fifth degree, if plotted upon coordinate paper, are sufficient to establish a
correction curve from which the error of any division upon the scale can be accurately found (see Fig. 83).
Verification
by Quartz Plates.
The
simplest and easiest

is
of scale verification, as well as the

fully standardized
most accurate,
quartz plates. The cost of to standardize the entire scale is, however, prohibitive, so that plates the chemist is usually content with a few standard plates for that
portion of the scale most used, as 80 to 100 for cane sugar.
method means of careby a sufficient number of
The
pos-
120
SUGAR ANALYSIS
session of a few carefully standardized quartz plates is a necessity for accurate saccharimetric work, not so much for standardization (since the constant error of the scale need be determined but once), but for
the determination of zero point, which observations.
is
necessary with each set of
The standard quartz plates furnished by instrument makers are mounted in metal tubes upon which is stamped the reading that the It is implates should give upon the particular saccharimeter scale. be verified this some that reading by testing bureau, as slight portant errors in marking or faults in optical homogeneity of the plate are not uncommon. The surface of the plate when placed in the instrument must be perpendicular to the beams of polarized light which traverse
it;
for this reason the plates should never
be loose in their mountings.
tightly upon the be errors in the as Rotation of might produced plate, optical quartz. the plate about the axis of its tube should cause no change in the field
On
the other hand, the mounting
must not press too
The plate when being used should be brought as at the end point. close to the analyzer diaphragm as possible in order to give the greatest
spread to the cone of light rays emanating from each part of the field. Care must be taken that the standard plate during polarization have
exactly the instrument.
it
same temperature as that of the quartz wedges of the If the plate have a temperature above that of the wedges,
a reading higher than
its,
will give
true value.
The temperature
polarization coefficient of quartz is 0.000136, so that the polarization

of a plate reading 100
V. at 20 C. would be for 30 C.,
100
(0.000136) (30
- 20) =
j
100.14
V.
If plate and instrument are of different temperature, the plate should remain several hours in the trough of the saccharimeter before using, that sufficient time may be given for it to acquire the same temperature. While it is necessary that quartz plate and wedge system have the same temperature, it is not essential that this be the standard temperature for the instrument, since the variations due to temperature
The slight differences are practically the same for plate as for wedge. due to effect of temperature upon shape of quartz wedge and upon
expansion of nickeline scale are expressed by the formula (Schonrock), Vw = V t V t 0.000005 (t - 20), in which F20 and V are the readings of the plate at 20 C. and t C. respectively. A standard plate polar-
izing 100
(plates
and wedges
V. at 20 C. would accordingly polarize 99.99 V. at 40 C. in each case at same temperature), a variation of

is
0.01
V. for 20 C. difference, which
negligible in practical work.
THEORY AND DESCRIPTION OF S AC CHARI METERS

Verification
121
by Pure Sucrose.
second means of verifying the
saccharimeter scale is with chemically pure sucrose. The preparation of sucrose of requisite purity is a matter of some difficulty; the method
of the International
sis
Commission
for Unifying
Methods
of
Sugar Analy-
is
as follows
purest commercial sugar is purified in the following manner: hot saturated aqueous solution, precipitate the sugar with a Prepare absolute ethyl alcohol, spin the sugar carefully in a small centrifugal machine, and wash in the latter with absolute alcohol. Redissolve
the sugar obtained in water, again precipitate the saturated solution with alcohol, and wash as above. Dry the second crop of crystals
"The
between blotting paper, and preserve in glass vessels for use. Determine the moisture still contained in the sugar and take this into account when weighing the sugar which is to be used." If a hand centrifugal is not available, the fine crystals of sugar may be filtered and washed In saturating the sugar solution free of sirup upon a Buchner funnel. before precipitation with alcohol, it is well not to heat above 80 C.
The sugar
solution thus prepared
is filtered
through a hot-water funnel
In this way the sugar is precipiinto the alcohol, stirring vigorously. tated in the form of fine crystals which are easily dried in the air.
Moisture is determined by drying at 105 C. In the selection of sugar for purification, the finest grades of small domino sugar (polarizing 99.90 to 99.95) have been found in the author's Rock-candy crystals, which are experience to give the best results.
sometimes recommended, should never be used; they frequently contain perceptible quantities of acid, with the result that inversion takes Complete absence of acidity in sugar and place during purification.
alcohol
necessary. verify the 100-degree point of the saccharimeter scale, the normal weight of sugar is weighed into a 100-c.c. flask, dissolved in dis-
is
To
tilled
water, and the solution made up to volume, care being taken that the liquid is well mixed before making up the last few cubic centimeters. The solution, which must be perfectly clear, is then polarized in a 200-
mm.
tube.
The
for the saccharimeter
conditions of weight, volume, and temperature required must be rigidly observed; the flasks and tubes em-
10 readployed should have been previously calibrated. The average of in the sugar, moisture the for corrected is result this taken and ings the amount of which must be determined in a separate portion with
each set of observations. The sugar used for polarization should not be dried in a heated-air or water bath owing to the danger of slight
*
1900. Proceedings of Paris Meeting, July 24,
122
SUGAR ANALYSIS
when changes in composition. If the vernier of the scale is set at the field is matched, the polarization of the sugar corrected for moisture should be exactly 100. In the same manner, other divisions of the
saccharimeter scale can be verified by taking fractions of the normal = 85-degree point of scale, etc.; see weight (e.g., normal weight X 0.85
Table XXI).
Verification
by Control Tube.
The most convenient means
of
verifying the scale divisions of a saccharimeter when using sucrose is by means of the Schmidt and Haensch control tube.* This method
presents the advantage that perfectly pure sucrose does not need to be used; in addition to this, but very few solutions are necessary for
verifying the entire scale. The control observation tube according to Landolt's latest form is shown in Fig. 82. It is telescopic in construction and can be adjusted
Fig. 82.
Control tube for verifying scales of saccharimeters.
so as to give a
column
of solution for
420
is
mm.
off
is
The
length of solution,
scale
read
upon the
any length between 220 mm. and is regulated by the screw T, S by means of the vernier J to 0.1 mm. The
which
surmounted by a funnel E, which does not serve for filling, but simply receives the overflow of solution as the tube is shortened. For filling the tube, the funnel is removed and the opening closed by means of a plug (P) the tube is then drawn out its full length and After rescrewing filled from the end by unscrewing one of the caps. the cap, the tube is set in an upright position and the funnel replaced as before. After shortening the tube slightly, a few cubic centimeters of solution are poured in the funnel, which is then closed with a small cap
tube
;
to prevent evaporation. In using the control tube, it is best to begin at the 100-degree point (which is supposed to have been previously verified) of the saccharim* Z. Instrument., 4, 169.

eter scale
123
and work downwards. A sugar solution is first made up of such concentration as to give a reading of 100 degrees at about 400 mm. This will be sufficient to test the scale the few divisions length of tube.
above 100 and all divisions below 100 to 55. If the reading, for example, is 100 at 400 mm. upon the tube scale, it should read 105 at 420 mm., 95 at 380 mm., etc. If a deviation be found at any division from the calculated value, other readings should be made at neighboring points
of the scale to determine the position of
maximum
error.
After test-
ing the scale to the 55th division (220 mm.), another solution must be prepared which will give a reading of 55 at about 400 mm. and the
scale tested
down
to 30.
By
proceeding in this way, always making
the final point of one series the starting point of the next, the scale can be tested its entire length with 5 solutions. Landolt* has given the
following table of concentration for solutions to be used with the control tube in testing the Ventzke scale:
Number.
124
SUGAR ANALYSIS
TABLE XXII
Verification of S.
&
H. Saccharimeter, No. 7075

No.
I
Series
Scale division of saccharimeter.

To
field,
125
both wedges are

screw.
verify the scales of a double-wedge saccharimeter, the scales of first set at zero with their verniers for the matched
of zero point being corrected by the regulating The working-wedge scale is then verified and its curve of error determined by the control tube in the manner described. The control
any deviation
then compared with the corrected readings of the working scale and its own error curve plotted. A still better direct method is
scale
is
100
95
90
85
80
75
70
65
126
tion obtained
SUGAR ANALYSIS
by the calculated weight of sugar is found to be 100, then the original scale reading of the saccharimeter is verified.
EFFECT OF TEMPERATURE UPON THE READING OF SACCHARIMETER SCALES

In the polarization of sugars and other materials upon quartz-wedge saccharimeters, the effect of temperature upon the scale reading is a most important factor. The saccharimeter is graduated to be used
at a fixed temperature (17.5 C. or 20 C.), and in the most carefully regulated sugar laboratories this temperature is maintained through-
out the year. But very few laboratories, however, are equipped with the necessary appliances for maintaining a temperature of 20 C. in summer, and the influence of temperature changes upon the saccharimetric readings and the methods for correcting the errors of the should therefore be considered.
same
Temperature Coefficient
of
Quartz.
The changes
in specific
rotation of sugars with variation in temperature are considered on These changes apply to measurements made upon any page 178.
kind of polariscope. But with the saccharimeter, as distinguished from the rotating polariscope, there must be considered an additional error due to the influence of temperature upon the quartz compensation of the instrument. This influence has been shown by Schonrock* to be
threefold.
There
is (1)
sion or contraction.
perpendicular to its 0.000007. The polarization value of the 100 point of the scale through change in shape of the wedge decreases with increasing temperature
the change in shape of the wedge by expancoefficient of expansion per 1 C. of quartz axis (rj) is 0.000013, and parallel to its axis (Y) is
The
by ??' -17, or by the coefficient -0.000006. There is (2) the change per millimeter thickness in the specific rotation of quartz itself, which for each degree increase in temperature increases by the coefficient
0.000136.
is
The combined temperature

There
is
coefficient of the
wedge system
the change due to the expansion and contraction of the material constituting the scale. The error due
therefore 0.000130.
(3)
to this change, together with that resulting from atmospheric humidity, was so great with the old ivory scales that the latter have been replaced
in
most saccharimeters with the alloy nickeline which has an expansion

1
coefficient per
C. of 0.000018.
The
total correction, therefore, for
a quartz-wedge saccharimeter with nickeline scale is 0.000148. The polarization value w for any temperature t is then expressed by the
*
127
= 0.000148 (t With saccharimeters whose 20) J. equation w* scale is etched directly upon the wedge itself, as is the case with Schmidt and Haensch instruments of recent construction, the coefficient remains
w\l+
0.000130.
The above increase in polarization of quartz with increase in temperature necessarily produces a lowering in the readings of the saccharimeter scale, since a smaller thickness of quartz is required for compensation.
With sugars which undergo a decrease in specific rotation with increase in temperature, the combined influences are in one direction and the With sucrose, for example, error thus introduced may be considerable. the temperature coefficient of polarization becomes at 10 C. 0.000390 (0.000148 + 0.000242), at 20 C. 0.000332 (0.000148 + 0.000184), and
at 30
of Sucrose. The variation in the Ventzke reading of the normal weight of pure sucrose for 1 C. change in temperature has been found by different authorities to be as follows:
Temperature
C. 0.000269 (0.000148 Coefficient
+ 0.000121).
Andrews* The United States Coast and Geodetic Survey

Wiley t
Prinsen Geerligs{.
0.0300
0.0293 0.0314
0.0300
0.0310
Watts
& Tempany
Average =
0.0303
T The average temperature
coefficient
of the
above
is
therefore
0.000303, which agrees with the figure of Schonrock for 25 C. (0.000148 = 0.000300. For temperatures between 20 and 30 C. the -f- 0.000152)
20 general equation F =F'Jl-f 0.0003 (-20){ may be used for changing the Ventzke reading (V ) of pure sucrose at any temperature t to the reading- (F20 ) at 20 C.
1
Temperature Coefficients
coefficients of
The temperature Other Sugars. other common sugars for readings upon the Ventzke scale
of
are given in the following table. The temperature coefficient for fructose and invert sugar are for readings made upon the negative scale of the
saccharimeter; while the coefficients of these sugars decrease the same as those of the dextrorotatory sugars, the direction of the decrease in both cases is towards the point and therefore opposite to each other
(as indicated
*
by the arrow
points).
Inst. Technology,
Technology Quarterly, Mass.
May
(1889), 367.
t J.
t
Am. Chem.
Soc., 21, 568.
Archief Java Suikerind, July (1903). West Indian Bull., Vol. Ill, p. 140.
128
SUGAR ANALYSIS
TABLE XXIII
Giving Temperature Coefficients of Different Sugars for Ventzke Scale
Sugar.

TABLE
129
XXIV
Showing Influence of Temperature upon Ventzke Reading of 1 per cent Sucrose, FrucSolutions made up to tose, and Invert Sugar for a Normal Weight of 26 gms. Volume at Temperature of Polarization
1
per cent sucrose per cent fructose
=
= =
-^ = -0.0003
^ = +0.0096
V. for
C. increase.
C. increase.
V. for
per cent invert sugar
^^ =
left.
+0.0048 V.
for 1
C. increase.
denotes change toward the
denotes change toward the right.)
Since the influence of temperature
upon the rotation
of glucose
is
so small as to be negligible, the change in rotation for 1 per cent invert sugar should be the same as that for 0.5 per cent fructose, or +0.0048 V. This is the result actually obtained, so that the calculation is verified.
SHALL SACCHARIMETERS BE ADJUSTED TO VARIABLE TEMPERATURES?
in
The International Commission* has provided that "for laboratories which temperatures are usually higher than 20 C., it is permissible
to graduate saccharimeters at
any suitable temperature, providing that the volume be completed and the polarization made at the same temperature."
The Commission has
neglected, however, to say
how
this
graduation shall be made.
It is evident that in order to
have a normal
weight of sucrose, under the conditions prescribed for a saccharimeter at 20 C., polarize 100 at 25 C. or 30 C., the compensating thickness of quartz in the wedge system must be made thinner for each part of the scale in order to counterbalance the decrease in specific rotation of
sucrose.
Owing, however, to the confusion and mistakes which would
arise
in the use of standard plates with saccharimeters of different compensating power, a better plan would be to make no change in the instru-
ment
but to alter the conditions of polarization, such, for example, as increasing the normal weight of sugar, or increasing the length of the observation tube, or decreasing the volume of the flask, any one of which means will bring the polarization of pure sucrose to 100 for any desired temperature above the standard. Since odd lengths of tube or volume of flask are undesirable as well as confusing, a change in the normal
itself,
Proceedings of Paris Meeting, July 24, 1900.
130
weight of sucrose
is
SUGAR ANALYSIS
the simplest of
all
means
of correction.
The method
*
of calculation can be understood
from the following example.
What would be
eter standardized at
the normal weight at 25 C. for a quartz-wedge saccharim20 C. for 26 gms. sucrose dissolved to 100 true c.c. and
coefficient of the specific rotation of sucrose at 22.5
polarized in a
200-mm. tube?
C.
The temperature
is
0.000168 (Schonrock).
is is
The temperature
coefficient of the nickeline scale
and quartz wedge

tion tube
0.000148; the expansion coefficient for the glass observa-
0.000008.
1
The new normal weight would then be
26,000
+(0.000148
c.c.
+ 0.000168 - 0.000008)
(25
dissolved to 100 true
in a flask standardized at 25
20) C.
26.040 gms.
saccharimeters are employed constantly in the investigation sucrose solutions, it might be advisable to make a change such as pure the above in the normal weight. But for varied work with different classes and mixtures of sugars whose specific rotations are affected in
of
When
opposite ways by changes in temperature, it is inaccurate to make alterations based upon the change in properties of one single sugar. The results obtained upon saccharimeters differently standardized are
then no longer comparable. The sucrose normal weight is frequently employed upon mixtures of sucrose with other sugars; in such cases changes in normal weight to correct for rotatory changes in the sucrose alone are wholly unwarranted. In view of the fact that the work of
saccharimeters is usually of a varied character, it seems best to leave the scale and standard conditions of the instrument unchanged. The chemist should work wherever possible under the conditions of tem-
perature prescribed for his saccharimeter, and when this cannot be done he should correct his readings as well as possible by a factor established for the particular product which is being examined. It must always be borne in mind that while the saccharimeter scale
established for the rotation of sucrose, its divisions indicate percentages only when pure sucrose is being polarized; in all other cases the
is
scale division
degrees polarization, degrees sugar scale, etc.) interpret according to his particular needs.
*
becomes a mere conventional number (degrees Ventzke, which the analyst must
This example is from a calculation supplied by the Physikalisch-Technischc Reichsanstalt, in reply to a suggestion by the author to use the old Mohr c.c. normal weight 26.048 gms. (17.5 C.) for true c.c. at 25 C. The old normal weight would give a reading of 100.031 V. when dissolved in 100 true c.c. in a flask standardized
at 25
C.
If
the true
c.c. flask
would be reduced to 100.019
V.,
standardized at 20 C. be used at 25 C., this error which is within the limits of error for observation.
THEORY AND DESCRIPTION OF SAC CHARI METERS

DESCRIPTION OF SACCHARIMETEBS
Tint Saccharimeters
131
in
The saccharimeter of Soleil as modified by Ventzke and Scheibler Germany and by Duboscq in France consists of an adaptation of the
quartz- wedge compensation to the polariscope of Robiquet (p. 86). The Soleil- Ventzke-Scheibler Saccharimeter. The construction
and arrangement of the optical parts in the Soleil saccharimeter as modified by Ventzke and Scheibler are shown in Fig. 84. A is a Nicol prism and B a plate of left or right rotating quartz cut perpendicular to its optical axis; these constitute the tint producer and are mounted
_,
F
Fig. 84.
LJ
Soleil-Ventzke-Scheibler tint saccharimeter.
in a
is
movable sleeve which can be rotated by a rod and pinion from J. a condensing lens, D the polarizer, and E a Soleil double quartz
plate (p. 86).
The quartz compensation is at F, the analyzer at G, and telescope at H. In using the instrument the telescope is focused upon the bi-quartz plate, so that the dividing line is sharply defined.
The
zero point of the scale
until both is then determined by turning have the same tint (in the manner described on p. 88). By rotating the regulator or tint producer from /, the tint which is most sensitive to the eye of the observer is obtained. This tint, which
sides of the field
is
different for different eyes, is usually of a

it will is
very delicate violet or
pearl color;
the Nicol
vary according to the angle with which In of the polarizer. set with reference to the Nicol
of course
order to remove the disturbances in transition tint due to colored

solutions
the adjustment of the regulator
(which cannot be remedied in the Robiquet polariscope), is changed until the tint is again of
is
With very dark solutions the transition tint greatest sensitiveness. almost a shadow owing to the absorption of color.
132
SUGAR ANALYSIS
as modified
differs
The Soleil saccharimeter The Soleil-Duboscq Saccharimeter. by Duboscq, the type of tint instrument used in France,
from the form previously described in that the Nicol producing
is
the sensitive tint
in Fig. 85. sensitive tint is produced with the quartz plate C,
by
situated in the eyepiece of the telescope, as shown until the The latter is rotated by a milled ring
instrument
telescope.
is
which in the Duboscq the objective of the and the situated between analyzer
telescope
is
The
focused upon the Soleil double plate at
Fig. 85.
Soleil-Duboscq tint saccharimeter.
D in or out; longitudinal guides prevent any which might disturb the tint. In the Duboscq instrument the two wedges of the compensator are of equal size, and, being
by moving the eyepiece
lateral rotation
driven past each other by the pinion in opposite directions, give a stratum of quartz of variable thickness. A scale and vernier, which follow the wedges in their movement, indicate the reading.
According to Landolt,* the average error of adjustment with the The instrument has 0.2 degree of the scale. saccharimeter is the same objection as the Robiquet polarimeter, in being unsuited to the color-blind. The adjustment of end point to color is also much
Soleil
more fatiguing to the eye than adjustment to uniformity of shade. Owing to these objections the color saccharimeter, although 20 years
ago the standard instrument,
use
is is
but
little
used at the present time.

of
Its
in fact
condemned by the Imperial Testing Bureau

Half-shadow Saccharimeters
Germany.
The various types of half-shadow saccharimeter used at the present time consist simply of an adjustment of the quartz-wedge compensation to some one of the half-shade polarizers previously described. The forms are the double-field saccharimeter with Jellet-Cornu principal
*
"
Das optische Drehungsvermogen"
(1898), p. 348.

polarizer; the double-, triple-,
133
Laurent plate;
and concentric-field saccharimeters with and the double- and triple-field instruments with
Lippich polarizer.
Saccharimeter with Jellet-Cornu Prism.
single-wedge halfis
shadow saccharimeter with Jellet-Cornu prism as

Fig. 86.
polarizer
shown
in
Fig. 86.
Single-wedge saccharimeter with Jellet-Cornu prism.
N. Sliding sleeve containing condensing lens. 0. Modified Jellet-Cornu prism (Schmidt and Haensch prism). E, F. Parts of quartz- wedge compensation. H. Analyzer. J. Telescope, which is focused upon the dividing line of the split prism at 0. K. Microscope for reading scale.
saccharimeter, which 15 years ago was the standard form employing the Ventzke scale, is at present almost entirely replaced with saccharimeters using the Lippich polarizer. Laurent's Saccharimeter. As a type of the saccharimeters conof instrument
The above
structed
in Fig.
87
by French instrument makers, the Laurent instrument shown is described. The arrangement of polarizer, half-wave plate,
and device for regulating the half-shadow angle is identical with that of the Laurent polarimeter (Fig. 72). The divided circle and rotating analyzer of the latter, however, are replaced in the saccharimeter by
the quartz-wedge compensation. The saccharimeter is adjusted to
until the
its
zero point
by
first
turning
If it should be found of the field agree in shade. that one side of the field has more of a reddish tinge than the other at the end point, the screw F, which controls the analyzer, is turned so as
two halves
to darken slightly the side of the field
most
colored.
The screw G
is
134
SUGAR ANALYSIS
if
then turned again to equality of shade;
there
is still
a difference in
again turned to equality of color, shade. By proceeding cautiously in this way the observer will at length reach the point where both sides of the field correspond in shade and When this point is reached the screw T is turned until the of color.
is
moved
slightly as before,
and
Fig. 87.
Laurent's single-wedge sacchari meter.
Lamp for producing white light (oil, gas, electricity, etc.), placed 200 mm. from B. B, E, R, K, J, X, U, D, L, the same as under Laurent polarimeter (Fig. 72). R. Saccharimeter scale, which with vernier V is illuminated by light reflected from
A.
A by the mirror N. Magnifying glass for reading scale and vernier. G. Screw for moving quartz wedges of the Soleil compensator.
.
the scale coincides with the
This adjustment should of the vernier. be verified by taking a number of check readings. The 100-degree point of the Laurent saccharimeter scale corresponds to a rotation of 21 40', the value given by French physicists to
the rotation of the 1-mm. plate of quartz. The normal weight for this angular displacement, as previously noted, is 16.29 gms. sucrose for
polarized in the 200-mm. tube. The Laurent saccharimmanufactured with a scale adapted to the so-called International normal weight of 20 gms. The instrument is provided with double or triple field, as desired. The scale divisions extend from to
100 true
c.c.
eter is also
110 to the right.

"Plaque Type."
is
135
rimeter
The 100-degree point of the Laurent sacchaa standard plate of quartz 1 mm. thick. This by " standard plate plaque type" also serves for the polarization of levoWith the plate in the trough of the instrument, solutions. rotatory the zero point of the scale is transferred to 100; levorotatory solutions
verified
are then simply read

difference
backwards upon the scale, the reading being the between readings of plate and solution. A solution, for
example, reading 67.4 with the 100-degree plate in position has a This method of polarizing levorotatory solu32.6. polarization of tions is of course applicable to all single-wedge saccharimeters. A 100-degree Laurent "plaque type " was remounted by the author and sent to the United States Bureau of Standards for a certification
as to its angular rotation
and its value in sugar degrees upon a saccharimeter employing the Ventzke scale. The rotation of the plate for sodium light of 589.23 wave length was given as +21.713
and the rotation in sugar degrees as read plate by the author upon a late-model Schmidt and Haensch saccharimeter gave a reading of +62.64, and
(T
20) =b 0.004,
+ 0.003
+62.65.
The same
upon a late-model Fric saccharimeter (Bates modification) a reading of +62.65. These readings of the " plaque type" not only prove the perfect identity of the Ventzke sugar scales employed by two
different
manufacturers, but also permit the establishment of the
exact ratio between the French
and German normal weights; for all other conditions as to the temperature and volume are the same in both these countries. The ratio 100 26 gms. :: 62.65 shows that
: :
the ratio of the

is is
to the French normal weight as 26 gms. to 16.289 gms., or, in even hundredths, 16.29 gms., which
German normal weight
normal weight prescribed in France. The Duboscq-Pellin saccharimDuboscq-Pellin Saccharimeter. eter for white light, as regards position of polarizer, half-wave plate,
quartz-wedge compensation, etc., is the same as that of the Laurent. The concentric field of the Pellin saccharimeter requires a somewhat
different cutting of the half-wave plate, but in other respects the two saccharimeters are very much alike.
identical with the official
The saccharimeter with Lippich polarizer is the form most generally The half-shadow angle between the prisms of preferred at present. the polarizer is usually between 5 and 8 degrees; it can be measured approximately by noting the interval between the points of maximum
light
The degrees Ventzke extinction each side of the zero point. between the two points of maximum darkness multiplied by 0.34657
gives the angle of the half shadow.
136
SUGAR ANALYSIS
A single- wedge Schmidt Schmidt and Haensch Saccharimeters. and Haensch saccharimeter upon tripod support with electric attachment for illumination is shown in Fig. 88.
Fig. 88.
Single-wedge Schmidt and Haensch saccharimeter with electric attach-
ment
for illumination.
V. Detachable end containing lamp and for inserting cell of bichromate solution. P. Position of Lippich polarizer for double or triple field. G. Casing of sheet brass for protecting wedges from dust.
eters
The method of scale illumination in Schmidt and Haensch saccharimis shown in Fig. 89 which gives the arrangements of parts for a double- wedge instrument. The light from the lamp is focused upon the small window a in the wedge housing, and is reflected from the mirror b through the ground-glass plate c upon the scale from which it is reflected through the prism p into the microscope whose objective is at d and eyepiece at / The working wedge is operated by the screw A g. and the control wedge by the screw K. The appearance of the scale of this instrument as viewed through the microscope is shown in Fig. 80. The latest and most improved type of Schmidt and Haensch saccharimeter is the double-wedge apparatus shown in Fig. 90. The instrument is mounted upon a bock or trestle support, and for saccharimeters which are in constant use this method of mounting is most satisThe factory as it insures perfect rigidity and accurate alignment. wedges are moved by milled screw heads at A and K which are so

H
137
m
:|
Fig. 89.
Device
for illuminating scale of
Schmidt and Haensch saccharimeter.
Fig. 90.
Double-wedge Schmidt and Haensch saccharimeter upon bock support.
138
SUGAR ANALYSIS
placed that the hand can rest upon the table during adjustment. The screw moving the control wedge can be fastened with a clamp, and is placed at a slightly higher elevation to prevent liability of confusion.
Peters's Saccharimeter.
Very similar
in construction to
the
above apparatus is the saccharimeter of Peters shown in Fig. 91. The long tube R prevents placing the light too close to the polarizer. The bichromate cell is placed within S; the cover C of the trough is not hinged but simply slides over or under the tube. The scale in the
Fig. 91.
Double-wedge Peters saccharimeter.
sheet-metal housing is illuminated by light reflected from the mirror L; a black paper disc P protects the eye against the glare of the obser-
vation lamp. Fric's Saccharimeter.

J. Fric
The half-shadow saccharimeters
of J.
and
are very similar in construction to the instruments previously described except in the method of scale illumination. In the latest
types of Fric saccharimeter a part of the light, as it passes from the source of illumination through the diaphragm at the end of the instrument, is reflected through a system of mirrors and lenses upon the scales. This illuminating attachment is shown in the Bates sac-
charimeter (L in Fig. 94), but the distinctive feature of the Fric illumi-

nating device
is
139
at the scale
The
light
from
is
reflected
end of the instrument as shown in Fig. 92. from the mirror A (which in the instru-
ments with enclosed wedges
is stationary) through the milk-glass plate the latter scale C, the in B upon the latest Fric saccharimeters being
made
of
glass.
The
light
from
L"
is
reflected
from the mirror
to the through the focusing lens eye of the observer. The divisions
of
the
scale
illuminated
in
this
manner appear with great

ness.
distinct-
The
Fric double-wedge in-
struments are provided with separate focusing lenses for reading the
working and control scales. The lens mountings and the milk-glass
plates for the two wedge systems are usually of different colors in
order to prevent confusion.
SACCHARIMETERS WITH VARIABLE SENSIBILITY

Of the instruments previously described, the French saccharimeters, using a Laurent half-wave plate and employing monochromatic or bichromate-filtered white light, are the only forms of apparatus which
permit a variation of the half-shadow angle to suit the requirements of
greatest sensibility.
In
is
all
fixed.
the Schmidt and Haensch saccharimeters the half-shadow angle An attachment for shifting the large prism of the Lippich
and regulating the half-shadow angle has been supplied by some manufacturers. While this regulating device presents certain advantages, it has been condemned by Landolt* on the ground that every change in the half shadow introduces a change in the zero point
polarizer
which has to be corrected by rotating the analyzer until the field is an impossible remedy in again evenly illuminated at the zero point a saccharimeter with fixed analyzer.
Bates's Saccharimeter. To obviate the objection last named, Bates f has devised an attachment which rotates the analyzer automatically and makes it possible to correct the zero-point error for any change
"
t
Das optische Drehungsvermogen,"

S.
U.
Bur. Stand. Bull., Vol.
4, p.
351. 461; Z. Ver. Deut. Zuckerind., 68, 105.
140
SUGAR ANALYSIS
The principle of in the half-shadow angle without resetting the scale. the Bates saccharimeter can be understood from Fig. 93. that Let OP be the direction of the plane of the large Nicol and
ON
a Lippich polarizer, let AZ be the plane of the analyzer at right angles to OB the bisection of the half-shadow angle PON or a. We will suppose for a moment that the intensities of light in OP and ON are equal and that the plane of the large Nicol be moved
of the small Nicol
.in
from
OP
to
OP' forming with the plane
of the small Nicol the
new
Fig. 93.
Illustrating principle of Bates's saccharimeter.
To obtain uniformity of field at the zero point for angle P'ON or a' the new angle a' the bisection OB must be moved to OB'. It will be
.
seen from the diagram that the angle
BOB' =
Zi
2i
~
2i
'
To
correct, therefore, for the displacement of zero point, assuming the intensities of light to be always the same for both Nicols, the plane of
the analyzer must be moved through one half the angular displacement of the large Nicol of the polarizer.
for the large
In the Lippich system, however, the intensities of light are not equal and small prisms of the polarizer. A part of the light is
141
extinguished in the small Nicol and there is also a loss from reflection and absorption. We will consider first the light lost by absorption.
Draw KL _L ON; amplitude of light from large Nicol. amplitude of light from small Nicol; the plane of the analyzer AZ must then be moved to A'Z' that the amplitudes OC and OF be equal in each half of the field. The angles AOA r and BOD, through which the plane of the analyzer and its perpendicular have
Let
then
OK =
OL =
moved,
is 5
of light in
OP
or the change from the true zero point when the intensities and are equal, in which case a = 0.
ON
suppose in order to increase the intensity of light for the half shadow that the plane OP of the large Nicol be moved to OP' inThe amplitude OK' remains the same as OK. Draw creasing a to a. K'U J_ ON; then the amplitude in ON = OU. The plane of the analyzer must now be moved to A"Z" in order that the -Is K'C' and
will
off the equal amplitudes OC' and OF' in the two halves of the OD' which is _L A"Z" will then form with OB', the bisection of The angle = DOD' through which the analyzer a', the new angle d'. has moved from its previous position is expressed by the equation
field.
We
L'F cut
In the polariscope of Bates (Fig. 94) the analyzing Nicol and the system are mounted in bearings and are The milled head, which operjoined by gears with a connecting rod.
large Nicol of the polarizing
ates the driving
mechanism,
is
shown
at
H.
When
the milled head
is
turned the two Nicols are rotated and the design of the gears is such that the analyzing Nicol always receives one half the angular displacement of the large Nicol of the polarizing system. Above the
milled head
is
position of the Nicols.
a circular scale which shows the polarizing angle for any In moving the plane of the large polarizing
Nicol through the angle POP' (Fig. 93) the rotating device of Bates's polariscope moves the plane of the analyzer through the angle BOB'. In this way the zero-point error of the instrument will always be equal
to the value of d for zero
any angle of the half shadow, assuming that the had been previously adjusted for a = 0. If the zero point of the instrument be set for any value of the half shadow a, and a be then
changed to
a',
the zero will have an error of

=
this value disappears
5'
d ( the
analyzer hav-
ing rotated
from the equation
142
SUGAR ANALYSIS
calculated values of 5 in Ventzke degrees for different values of a according to the two equations,
The
the half-shadow angle
tan
tan 3 s 2
and
tan
1 =1
COS
a V0.92
,
a
tan 2
+ cos a V0.92
(see p. 96), are
given in the following table.
TABLE
XXV
Giving Calculated Values of Error in Zero Point for Bates 's Saccharimeter
VALUES OF
IN
VENTZKE DEGREES.

omission
143
may
occasion no serious error in the polarization of colored
solutions (as of low-grade sugar-house products), a bichromate light filter is required in the examination of high-grade cane sugars, starch-
conversion products, and many other substances. An absorption cell for this purpose should be placed just in front of the aperture between
the saccharimeter and the source of light. A very commendable feature of the Bates instrument is the thermometer (T Fig. 94) which indicates
the temperature of the quartz wedges.
Fig. 94.
TF, milled
Bates saccharimeter with variable

for operating
sensibility.
head
working wedge.
C, milled head for operating control wedge.
w, microscope for reading working wedge scale. c, microscope for reading control wedge scale. " " degrees of brightness or half-shadow angle. S,_ scale indicating
SACCHARIMETERS WITH MAGNIFIED SCALE.

For special kinds of work involving the investigation of products with a narrow range in composition, saccharimeters have been constructed with a limited magnified scale.
The saccharimeter devised

an
by Stammer,* shown
*
in Fig. 95, for polarization of sugar beets is

144
SUGAR ANALYSIS
example of such an instrument. In this apparatus a magnified scale, to 35, is attached to the side of the instrument at the reading from observer's left and permits the reading of polarizations with the unaided
eye.
The
is
which
wedge.
pointer of the scale P is moved by the tension roller R, connected by a small steel chain with the movable quartz
by
adjust the saccharimeter the field is brought to a uniform shade when the of the wedge scale and vernier at Q should turning
To
coincide.
If
the latter
is
not the case, coincidence
is
affected
by turn-
Fig. 95.
Stammer's saccharimeter with magnified scale for polarizing sugar beets.
should mark ing the regulating key V. In this position the pointer the S. there be any zero division of the scale Should exactly large
deviation the error is corrected by turning the adjusting lever L until the pointer is exactly at 0. Turning the screw to any division upon the wedge scale Q should then give the same reading upon the scale S. If this is not the case the error is corrected by turning a small
control screw
roller.
upon R which increases or diminishes the diameter of the The adjustment is one which requires considerable care and
should be checked repeatedly. Saccharimeters of the above type are especially adapted for the polarization of mother beets for seed production; they are constructed for tubes of 200 mm., 400 mm., and 600 mm. length.
Similar to the above instruments for sugar-beet analysis, saccharim-

eters
145
have been constructed with a magnified scale reading between 80 and 100 for polarization of sugars. These are manufactured usually only for use with tubes 400 mm. long, and employ a normal weight of
26 gms. to 100
c.c. solution.
necessitates of course doubling the interval
Doubling the length of observation tube between the scale divisions
and thus facilitates the reading. Instruments with a magnified limited scale will be found to relieve eye fatigue, where large numbers of analyses of a single product have With one person to prepare the tubes of sugar to be performed. solutions, a second to manipulate the saccharimeter, and a third to note the readings, a large number of polarizations can be made in a very
short period of time.
CONVERSION FACTORS FOR POLARISCOPE AND SACCHARIMETER SCALES

In the following table factors are given for converting 1 degree of the various polariscope scales into its equivalent in circular degrees, The conversions or in degrees of the different saccharimetric scales.
are based so far as possible
upon recent information supplied by the

Equivalent.
manufacturers of the several instruments.

Scale.
= 0.34657 angular rotation D. 1 angular rotation D = 2.88542 Ventzke sugar scale. = 0.21666 angular rotation D. 1 French sugar scale 1 angular rotation D = 4.61553 French sugar scale. = 0.62516 Ventzke sugar scale. 1 French sugar scale 1 Ventzke sugar scale = 1.59960 French sugar scale. = 0. 13284 angular rotation D, 1 Wild sugar scale 1 angular rotation D = 7.52814 Wild sugar scale. = 0.38329 Ventzke sugar scale. 1 Wild sugar scale 1 Ventzke sugar scale = 2.60903 Wild sugar scale. = 0.61313 French sugar scale. 1 Wild sugar scale 1 French sugar scale = 1 63098 Wild sugar scale. (Normal weight = 26. 00 gms. Ventzke scale; 16. 29 gms. French scale;
1
Ventzke sugar scale
10. 00
gms.
Wild
scale.)
scale is employed upon the Schmidt and Haensch, and Fric saccharimeters. The French sugar scale is employed Peters, the Laurent-Jobin and upon Duboscq-Pellin saccharimeters. The slight differences in ratio between normal weights and scale equivalents have already been discussed.
The Ventzke sugar
CHAPTER
VII
ILLUMINATION OF POLARISCOPES
the illumination of polariscopes and saccharimeters numerous been devised and the chemist must be guided in his selechave lamps tion by type of instrument, nature of substance to be polarized, and the kind of light supply available. Before describing the various types of lamps, a word should be said regarding the general subject
of illumination.
FOR
neglected point in the illumination of polariscopes and saQcharimeters is the placing of the light at the proper distance from the condensing lens. The light should never be placed so near as to over-heat the metal at the end of the instrument; neglect of this precaution may cause a softening of the balsam and wax mountings of
the polarizer and lead to serious derangement of the optical parts. The proper rule in setting up the polariscope is to place the light in
A much
such a position that
its
image
is
clearly defined
upon the analyzer
accomplished by fastening a needle or other sharp-pointed object just before the light and moving the instrument or light until a clear inverted image of the point is obtained upon a
diaphragm;
this is best
When piece of white paper placed before the analyzer diaphragm. the light is thus focused the polariscope is least susceptible to changes in zero point. The proper position of polariscope with reference to
light
can be seen from Fig. 96, which shows the arrangement of the When correctly placed optical parts in a double-wedge saccharimeter. an inverted magnified image of the light 7 is obtained at A. The
reciprocal of the focal distance of the condensing lens will then equal the sum of the reciprocals of the distances of lens from light and of
lens
from image.
In the case of a Schmidt and Haensch saccharimeter the focal
Example.
distance of the condensing lens was found to be 5 inches; the distance from lens to analyzer diaphragm was 20 inches; the distance for placing the light
would then
= -or6f be--h^: X ZO O
inches from the condensing lens.
146
The
telescope
147
(Fig.
96)
is
dividing line of
to the point of
the
field at
C and
focused by the observer upon the the analyzer or compensator turned
even illumination. The dividing line at C will then entire field appear of equal intensity. the and This will be disappear the case even with slight variations in intensity in different parts of the illumination, since at the point C, upon which the eye of the observer is focused, the light from any part p of the illumination will be dispersed through different parts of the field (as shown in the figure by
the dotted lines); any slight uneveness in the source of illumination Great irregularwill thus be distributed and not noticed by the eye.
ities
in illumination, however,
must be avoided, and
for this reason
it
JP
Fig. 96.
is
Showing method
of illuminating polariscopes.
important that the instrument be kept in perfect alignment with

longitudinal axis at right angle to the source of light.
fixed.
its
It is best to
Polariscopes mounted upon trestle supports are preferable to those upon tripods since a slight knock may swing the latter out of alignment and cause a change in
have instrument and light rigidly
the zero point. Variations in the brightness of illumination are also undesirable and for accurate work the emission of light should be constant. The
optical
589.22
center of gravity of purified sodium light, for example, is variations in this HJJL for a certain average brightness of flame;
brightness, however, may change the wave length by 0.11 ///* with corresponding differences in the rotation of polarized light (25" for a
rotation angle of 20 degrees).
With
salts of the alkalies
and
alkaline
earths, increasing the brightness of flame (increase of vaporized salt per unit volume of flame) produces an irregular broadening of the
spectral lines with a shifting of the nd of the spectrum.
mean wave
length toward the red
Of the various polariscope lamps for Light. few the more common forms will be described. a of light only The lamp shown in Fig. 97 illustrates the essential principles of most sodium lamps. This consists of a Bunsen burner with side entrance
Lamps
for
Sodium
sodium
for gas at s to
salt;
prevent stoppage of inlet through dropping of fused the burner is surmounted by a chimney which can be adjusted
148
to the desired height
SUGAR ANALYSIS
by the screw
h.
The holder
for the fused salt
bundle of fine platinum wires attached to an upright support and can be moved in and out the flame through a slot in the chimney by means of the screw p; the door k, which closes the front of the chimney,
consists of a spoon-shaped
shine through
allows only the brightest section of the flame to and excludes
the greater part of the heat. The flame is adjusted so as to
be
colorless,
with as strong an
free
air blast as possible, that the
light
may be
from incan-
descent carbon particles. In place of wire holders for
the salt
many
sodium lamps use spoons or V-shaped boats of sheet platinum or nickel, which are in some cases perforated with fine openings. The hot part of the
flame
impinges
upon the spoon and

produces a sheet of
Fig. 97.
Simple form sodium lamp.
of
sodium
each
light
upon
Fig. 98.
Pribram's
side.
The
sodium lamp.
must be renewed as fast as vaporized; a convenient means * is shown in Pribram's sodium lamp, Fig. 98, which contains two boats; the empty one is drawn out for refilling and
fused salt
of effecting this renewal
the one in reserve inserted in
its place.
f, Fig. 76, gives a more intense flame than either of the lamps just described. It consists of a powerful Muencke gas burner with cylindrical chimney L. Upon the latter are
The sodium lamp
of Landolt
placed two heavy nickel wires supporting rolls of fine nickel wire netThe burner is surmounted by a second ting which contains fused salt.
rectangular chimney of sheet iron with a movable brass door containing apertures of 20, 15, and 10 mm. diameter.
The simplest and cleanest of sodium lamps and the one giving the most continuous flame is that of Zeiss, Fig. 99. This is composed of
*
Z. analyt.
Chem., 34, 166.
f Z.
Instrument., 4, 390.
149
an upper part A, capping an ordinary Bunsen burner and secured to The casting A carries the it by means of a screw. of which the rectangular out diaphragm-screen K, the flat burner C producing a L is also cut, opening
square flame, and a small support for the salt carrier
E, which consists of a piece of pumice stone, measuring about 4
upon the
regulated It is best to adjust operating on the spring GH. the pumice stone so that it merely touches and
tinges the flame. flame, the latter
If
is
X 1 X J cm., saturated with salt. It is held support by the spring clip F and can be to the flame by means of the screw /
E be too deeply inserted in the over-cooled and a dark, rather

is
sharply defined zone
produced.
The
flickering
margins of the flame are cut off by the diaphragm K. A few minutes are needed for heating the pumice before the flame attains its maximum brilliancy,
after
which
it will
The
tablets of
remain constant for hours together. pumice stone saturated with salt are
at small cost.
salt,
supplied
by the trade
In place of
common
sodium bromide
is
sometimes used for illumination. This gives a much stronger flame, but the vaporization is much more rapid than with salt and there is the additional
Fig. 99.
The
Zeiss
may
disadvantage of giving off bromine vapors which attack the instrument unless the lamp is placed under a hood. Sodium carbonate, sodium phosphate, sodium nitrite, and mixtures
of these
with salt in various proportions are also used for sodium

Sticks of fused sodium carbonate heated hi
lamps.
an oxygen blast lamp give a flame of great brilliancy, and this is the form of light recommended by Landolt * when intense illumination is desired. Purification of Sodium Light. For accurate polariscope measurements it is necessary to purify the sodium light from other rays. This can be done either by use of light filters or by spectral separation of
the extraneous rays.
violet
Sodium end
light
of the
can be freed from most of the foreign rays at the spectrum by means of bichromate solution, which
has a strong absorption band in the green and blue. The rays at the other end of the spectrum can be removed by uranous sulphate solution,
which has a strong absorption band in the red. * "Das optische Drehungsvermogen " (1898),
combination of
p. 359.
150
these
SUGAR ANALYSIS
two
solutions, as in the Lippich light filter, constitutes the
most
effective
absorbent means
Lippich Light Filter. cell closed at the ends by tightly fitting cover glasses and divided by a The larger cell, 10 cm. glass plate into two smaller cells of unequal size.
long, is filled with
of sodium-light purification known. The Lippich light filter consists of a tubular
the smaller
cell is filled
a 6 per cent filtered solution of potassium bichromate, with a solution of uranous sulphate, 11(804)2,
3 2 0, prepared as follows: 5 gms. of purest uranyl sulphate, UO 2 S04 2 of and in 100 c.c. of dissolved are powdered chemically water, gms. pure zinc added; 3 c.c. of concentrated sulphuric acid are then added
in 1 c.c. portions, waiting each time until the evolution of hydrogen has nearly ceased; the flask is corked during the reaction, and is allowed to stand about six hours, when the solution is filtered and the cell imme-
+ H
diately filled in such a way as to leave only the smallest possible bubble After standing for a day the cell is ready for use; the of air behind.
uranous solution retains

its
its stability for
one to two months, or until
deep green color
is
changed by
oxidation into the yellow of the
cell must be refilled with fresh solution. The weights and volumes prescribed for making up the absorbent solutions must be rigidly adhered to. The spectrum purification of sodium light by means of glass prisms is the most thorough of all methods of purification. The process, which is a somewhat complicated one, is required, however, for only the
uranyl compound,
when the
finest
physical measurements.
for
wave lengths
length of the
sodium
line is
light
Landolt gives the following average from different sources in which the wave
placed at 589.62
w and the A
Purification.
line at 589.02
^.
TABLE
Wave Length
Number.
Source
XXVI
Kinds of Sodium Light
Wave
in
of Different
of light.
length
/x/x.
Bunsen flame with NaBr
Bunsen flame with NaCl..
Burner with NaCl or NaBr.

j
K Cr O in water. 10 cm. layer of 9 per K Cr O in water. Lippich filter K Cr O

2 2 7
10 cm. layer of 9 per cent
592.04 589.48 589.32

589.25
cent
7
and
U(S0
4) 2.
Perfectly
spectral
pure;
Sodium
light
<
light of only the
two
Landolt lamp with NaCl
lines. 1.5 cm. layer of 6 per cent 2 Cr 2 7 in water.
10 cm. layer of 9 per cent]
Bunsen flame with NaCl

Landolt lamp with NaCl
<
K O K Cr O
2
2
588.94
cm. layer
in water and 1 of 13.6 per cent
588.91
j
CuCl 2
in
water.
Unpurified
588.06
The Lippich
light filter gives a
151
wave length exactly between the and two D lines of sodium agreeing very closely with that obtained by In all cases where light filters are used the soluspectral purification. between be tions must lamp and condensing lens (see Fig. 96). placed For illuminating polariscopes and sacWhite Light. Lamps for a white with charimeters large number of lamps have been devised light, for use with oil, alcohol, gas, acetylene, and electricity.
100.
Hinks's
oil
lamp
Fig. 101.
with duplex burner.
Hinks's gas lamp with triplex burner.
convenient form of
is
oil
support burner is shown in Fig. 101. The metal chimneys of these lamps are usually fitted on the inside with a porcelain reflector; the focusing lens which is sometimes placed in the aperture of the chimney should be
that of Hinks, Fig. 100.
lamp with duplex burner and adjustable The Hinks gas lamp with triplex
removed as
it
may
cause an incorrect passage of the beams of light
through the polariscope.
152
SUGAR ANALYSIS
The best forms of gas lamp for illuminating are those provided with an Auer or Welsbach mantle (Fig. 102). The outer cylinder of these lamps, composed of sheet metal or asbestos, contains an opening whose lower half is covered with a plate of ground glass for diffusing the light; the upper uncovered part of the opening serves for illuminating the polariscope scale. A form of lamp for burning alcohol somewhat Gas burners for similar in design to the above is shown in Fig. 103.
producing lime or zircon light are also used for illuminating polariAcetylene lamps of 25 to 50 candle power give a light of great scopes.
Fig. 102.
Gas lamp
Fig. 103.
with Welsbach mantle.
Alcohol lamp with Welsbach mantle.
Fig. 104.
Stereopticon
electric
lamp.
brilliancy
and are
is
especially valuable
upon sugar plantations where gas
or electricity
acetylene lamps should be fitted with cylinders similar to those in Figs. 100 or 102. For electrical illumination a stereopticon 32-candle-power incandescent lamp is very suitable (Fig. 104); the closely wound filament concentrates the light within narrow compass, giving great intensity of
not available.
The
illumination. These lamps are best mounted in cylinders similar to that in Fig. 102; a plate of ground, glass is necessary for diffusing the
light,
153
suffi-
otherwise the irregularities in source of emission will not be

small electric attachment devised
is
ciently equalized for obtaining a uniform field.
illuminating their saccharimeters
by Schmidt and Haensch for shown in Figs. 88 and 105. The
Fig. 105.
Schmidt and Haensch
six-volt saccharimeter
lamp.
small lamps are adapted for a six-volt current which is supplied by a The storage battery or from the main line after reducing the voltage. is is screwed on which controlled S the switch apparatus (Fig. 88) by
the polarizing end of the saccharimeter. The electric lamp is held in two in connection with the two which are position by spring clips
The lenses 2 and KI (Fig. 105) project the light upon the the analyzer. As the horizontal filament is not always of diaphragm concentric to the frame, a vertical adjustment is necessary. To quite
terminals.
work the adjustment, the ring Z), which carries the by the screw and projecting arm 6. If the lamp is
lens
2 , is
rotated
also to be used for
illuminating the scale of the instrument, the mirror S' (Fig. 88) is set at an angle of 45 degrees, in which position the reflected light is concentrated by the lens upon the opening a (Fig. 89).
POLARISCOPE TUBES
For retaining sugar solutions during polarization there are a variety
In the selection of tubes of different construction, form, and length. the nature of his less of these the chemist must be guided more or by
work.
All tubes, however,
when accuracy
of observation
is
desired,
must conform to three general requirements: (1) the length of the tube must be accurately fixed; (2) the ends of the tube and the surfaces of its cover glasses must be plane parallel; (3) the tube must be centered evenly in its mountings and, when fitted with its caps, should
be free from eccentricity.
There are other minor requirements of
154
SUGAR ANALYSIS
tube construction which will be given under the description of the

different forms.
most common and simplest forms of glass polarand other forms of tube are usually supplied in lengths of 25 mm., 50 mm., 100 mm., 110 mm., 200 mm., 220 mm., 400 mm., 500 mm., and GOO mm.; for special kinds of work tubes of several meters' length have been constructed. A tube of 200 mm. length is used for the normal weight of all saccharimeters. If, on account of depth of color, a 100-mm. or 50-mm. tube is employed and the resultant reading is recalculated by mulFig. 106 shows the These ization tubes.
tiplying
by 2 or 4, there is, of course, a corresponding doubling or quadrupling of the errors of observation; short observation tubes are to be used therefore only in extreme cases. With very dilute sugar solutions
25
mm.
tube.
100
mm.
tube.
200
Fig. 106.
mm.
tube.
Forms
of plain glass polariscope tubes.
and with sugars or sugar mixtures of low specific rotation the 400-mm. or 600-mm. tube will increase the accuracy of the observation, provided the color be not too great to disturb the reading. Tubes of odd lengths, such as 55 mm., 110 mm., 220 mm., etc., should be distinctly marked lest they be confused with the 50-mm., 100-mm., and 200-mm. sizes. The ends of the glass observaMounting of Polariscope Tubes. tion tubes are cemented into metal mounts which are threaded for the purpose of receiving the screw cap. Litharge and glycerine make a much better cement than the waxy material employed by most manufacturers. The latter substance, especially on warm days, softens readily and when in this condition there is danger in screwing on the cap of drawing the mount from its setting so that it projects slightly beyond the ends of the tube; the length of the column of liquid to be polarized may thus be increased and a considerable plus error introduced
in the observation.
The ends
only slightly beyond the threaded heads; if too exposed there is danger of chipping or breakage.
of the glass tubes should project much of the end is
The chemist should not attempt to reset his tubes unless he has a small lathe in which they
can be centered and revolved while the cement may not be evenly mounted.
is
155
hardening, otherwise
the tubes
simple means of testing for eccentricity of mounting to place the tube, with caps screwed on, in the trough of a polariscope and while giving it a rotatory motion to
is
view the opening through the tube with reference to the If the tube has been properly centered polariscope field. and the caps are free from eccentricity the tube opening will
remain
in the center of the field
and show no wabbling
movement during
rotation.
To
test for plane parallelism of
the ends of the tube and of cover glasses, the experiment just described is repeated with the cover glasses in position
and the tube
filled
with water.
If
the ends of the tube have
not been ground squarely across or the cover glasses are not plane parallel, the opening of the tube will wabble perceptibly during rotation owing to the refraction of light through the water from the inclined surfaces of the cover glasses. A
Ventzke degree may be noted between the readings of a tube in different positions through lack of plane parallelism in ends or cover glasses. According to Landolt the angle between the opposite ground-end surfaces of a polariscope tube should always be less than 10 minutes and the angle between the two planes of a cover The small angles of inclination glass less than 5 minutes. between planes of cover glasses and between ends of tubes not exceeding 200 mm. in length is measured by a spectrometer. Calibration of Polariscope Tubes. A most convenient
difference of several tenths of a
means
of calibrating the length of polariscope tubes is the measuring gauge of Landolt, shown in Fig. 107. This gauge, which has an adjustable handle c, consists of a measuring
steel graduated for a distance of 400 mm. and with a sliding vernier b which gives readings to provided 0.1 mm. The lower end of the rod and the bottom of the
rod
of
vernier are provided with knife edges. When the knife edge of the rod is placed upon a smooth hard surface, such as glass, and the vernier brought down until its knife edges are in Fig. 107.
with the same surface, the zero point of scale gauge f or If there is lack of agreement, the agree. calibrating zero point of the vernier may be either adjusted or the differ- polariscope tubes, ence noted and applied to all readings. To calibrate an observation tube, one end of the tube is closed with its cover glass and
close contact
and vernier should
156
cap,
SUGAR ANALYSIS
and
after placing in
is
the measuring rod

glass; holding the
an upright position with the closed end down inserted until its knife edgejtouches the cover
rod perfectly upright the vernier is slipped down with the upper end of the tube; the readvernier will the and then give the length of tube. Other of scale ing the rod a little each time from its original are made, rotating readings
until its knife edges coincide
position,
and the average taken.
Calibration of tubes should be
made
at the standard temperature 20 C.; if measurements are made at temperatures very different from this the changes in length of tube and gauge due to expansion or contraction must be taken into account (coefficient of
expansion in length
C. for steel
0.000008). Measuring gauges can be verified as to accuracy at the Government Bureau of Standards. The measuring gauge of Landolt will detect an error of 0.1 mm., which is equivalent to an error of 0.05 V. for a sugar solution polarizing 100 V. in a 200-mm. tube. This is sufficiently close for ordinary saccharimetric measurements; if a finer determination of tube length is desired the measurement must be made upon a comparator; by means of this instrument measurements can be made to 0.01 mm. Cover Glasses. The cover glasses used upon polariscope tubes must be of strong, colorless, and optically inactive glass; their surfaces must be plane parallel and free from cracks or scratches. In screwing
0.000013 and for glass
the caps upon observation tubes, care must be taken that no severe pressure is brought to bear upon the cover glasses; otherwise the strain will render the glass optically active and produce serious errors in the
If a cover glass is optically active turning the tube in the trough of the polariscope will usually show variations in the intensity of the field with considerable difference in the reading for various positions of the tube. The practice of rotating the observation tube be-
observation.
tween readings is always a good one; in this way errors due to defective cover glasses, bad washers, pressure of caps, eccentricity, etc., may be detected which would otherwise escape notice. Cover glasses which have been rendered optically active through pressure should not be
used for a day, in order that sufficient time to neutrality.
may
elapse for readjustment
Washers. Another common source of error in polariscopic are badly fitting rubber washers in the screw caps of the tubes.
work The
washers should be of soft rubber and lie evenly against the back of the cap without the slightest marginal elevation, otherwise the washer
in tightening the cap
may give the cover glass an inclined position and cause a considerable increase in the reading.
Special
157
Forms
of Polariscope
Tubes
Schmidt and Haensch Tube with Enlarged End. Another form of glass polarization tube which presents several advantages is the Schmidt
and Haensch tube with one end enlarged (Fig. 108). The enlargement serves as a receptacle for any air bubbles which may be enclosed with
the liquid; the retention of a small air bubble in the tube is in fact desirable since, by moving the bubble through the liquid from end to end
Fig. 108.
Schmidt and Haensch polariscope tube with enlarged end.
(Air bubbles are collected at the point a, outside of the field of vision.)
before reading slight differences in temperature are equalized, and no troublesome striations, due to currents of solution of different tem-
Tubes without enlargement perature, are present to distort the field. must not retain air bubbles with the liquid; if striations are present
the tube
must remain at The most frequent cause

in the
rest until the solution has reached equilibrium.
is the warming of the solution tube by the hand; for this reason tubes should be handled only by the metal caps when placing in the instrument.
of a striated field
Fig. 109.
(a)
200
mm.
(6)
Landolt polariscope tube with sliding cap and enlarged end; 200 mm. metal polariscope tube.
To prevent the liability of excessive pressure cover Landolt has devised a tube with sliding cap, upon glasses, which is shoved into position over the metal mount (Fig. 109a). The
Landolt' s Tube.
French manufacturers also provide a cap that is shoved on and fastened with a bayonet catch. Tubes with screw caps, however, are the ones most preferred and, if care be taken not to draw them up too tightly, will be found to answer all requirements. When observation tubes are used in large numbers it is a great advantage to have
all
caps interchangeable. Metal Polarization Tubes.
Polarization tubes of brass or nickel or
other metal are preferred
by many
which
is
shown
in Fig. 109b,
Such tubes, a form of chemists. have the advantage of greater durability,
158.
SUGAR ANALYSIS
in the
but the disadvantage of being susceptible to the attack of acids (as method of inversion) or other corrosive liquids. Brass tubes
coefficient of
have also more than twice the
expansion of glass tubes,
Fig. 110.
Pellet's
tube for continuous polarization.
the coefficient
brass.
(/?)
for 1
C. being 0.000008 for glass and 0.000019 for
For glass and brass tubes measuring exactly 200 mm. at 20 C., the length at 35 C. (Le = L20 [l +0 (*-20)]) = 200.024 mm. for glass
i
Fig. 111.
Glass polarization tube with metal jacket.
and 200.057 mm. for brass, errors in length of no great significance. A more serious objection against metal tubes is the danger of their being bent out of alignment through hard or long usage. A knock or fall may cause a metal tube no apparent injury yet may bend it sufficiently
to produce a considerable error in the number polariscope reading. of brass polariscope tubes, recently submitted to the author for examina-
tion,
159
of the polariscope caused
were so badly out of alignment that rotating the tubes in the trough a difference of over 0.2 V. in the reading. Pellet's Tube for Continuous Polarization. In the polarization of a
large
number
of solutions in succes-
the analysis of sugar beets, juices, etc., the Pellet tube for continuous polarizations is often of great
sion, as in
use.
made
110.
Sections of this tube, which is of metal, are shown in Fig.
The ends of the tube
are closed
and
after placing in the instrument
the solution to be polarized is poured through a small funnel into one of the nipples, a or
ing through
6,
the excess escap-
an
exit
tube connected
to the nipple at the opposite end. As soon as the solution is polarized, the succeeding
solution is poured into the tube; the disappearance of striations and the clearing of the field indicate when
by rubber tubing
the previous solution has been

pletely displaced.
will
com-
The
Pellet tube
accomplish a valuable saving of time in certain kinds of work, but it is usually advisable to limit its use
to sugar solutions of approximately the same density; to displace a concentrated sugar solution with one
that
is
versa,
is
exceedingly dilute, or vice attended with more or less
risk of error.
Jacket.
Tube with Metal For polarizing sugar soluReservoir for supplying Fig. 112. tions, where the temperature must water of constant temperature. be measured or controlled, a jacketed observation tube such as shown in Fig. Ill is recommended. This consists of an inner tube of glass or metal with a central opening, c, which can be used for filling and for inserting a thermometer; an outer mantle of brass or nickel surrounds the inner tube and is provided with nipples for inlet and exit of hot or cold water as may be desired.
Polarization
160
SUGAR ANALYSIS
For supplying water of constant temperature for observation tubes, the Zeiss apparatus described on page 59 may be used. A form of water supply reservoir with stirrer, recommended by Landolt,* is shown The reservoir, which is insulated, is filled through the in Fig. 112. D. opening A with water to the desired level, indicated by the tube desired the burner to a temperature, The water is heated by means of shown by the thermometers at C, the heat being equalized by raising
and lowering the stirrer B. A form of constant temperature bath designed by Hudson f is shown in Fig. 113. The mechanical stirrer not only secures an even temperature through the bath, but also acts as a rotary pump which
From.Polarimeter
Mercury
Sealed Joint
To Polarimeter
\
Fig. 113.
Hudson's constant temperature water-bath.
creates a constant circulation of water as
shown by the
direction of
the arrows.
When solutions are polarized at temWiley's Desiccating Caps. peratures below the dew point of the atmosphere, the cover glasses of the observation tube must be protected against condensation of moisture
by means
of desiccating caps such as designed by Wiley J (Fig. 114). These are generally made of some non-conducting material such as hard rubber: they are closed at the end with a tightly fitting cover glass and contain a tube for holding calcium chloride or other desiccat-
ing substance.
"Das optische Drehungsvermogen " (1898), pp. 397. Am. Chem. Soc. 30, 1572. Hudson, J J. Am. Chem. Soc.
J.
18, 81.
161
When solutions are polarized at very high temperatures as at 87 C. (the point of inactivity for invert sugar) the use of glass, unless carefully annealed, for the inner tube of the water jacket is precluded.
Polariscopic
work
at
high
temperature
is
generally
performed in
(I)
Fig. 114.
(I) Threaded cap of polariscope tube. (II) Dessicating cap which screws on over threads of (I) t, removable glass tube containing dessicating substance s; w, inner perforated metal tube; g, cover glass held in position by threaded disk r; the disk is unscrewed by inserting a spanner in the two holes marked in black.
;
jacketed tubes constructed entirely of brass or nickel, the inner surface of which has been gold plated. The length of a 200-mm. tube (20 C.) at 87 C would be 200.107 mm. for glass and 200.255 mm. for brass,
equivalent to a plus error of 0.054 V. and 0.128 V. respectively for solutions polarizing 100 V. in a 200-mm. tube.
Fig. 115
Yoder's volumetric polariscope tube.
tube
volumetric polariscope Yoder's Volumetric Polariscope Tube. tube of is convenient for certain kinds of saccharimetric work.
this description, designed
by Yoder,
is
shown
in Fig. 115.
The capacity
of the tube to the graduation
mark upon the neck
is
10 c.c. By varying the length and diameter the tubes can be adjusted to any convenient volume.
162
SUGAR ANALYSIS
BALANCES FOR POLARISCOPIC
WORK
For the operations of weighing in saccharimetric work three types of balances are required, an analytical balance, a so-called sugar balance, and a balance for coarse weighing. The analytical balance should have a capacity of 200 gms. and with this load be sensitive to 0.1 mg. Such a balance is required for all
analytical processes, for determination of specific rotations, for calibration of flasks, weighing of pycnometers, and all other operations
where accuracy will answer for
is essential.
this purpose.
balance of the type shown in Fig. 17 With this balance a set of accurate
analytical weights (including one 100 gms. weight) will be needed.
Fig. 116.
Sugar balance.
In addition to the above a less delicate balance, sensitive to 2.5 mgs. with a load of 250 gms., will be required for the rapid weighing of definite
amounts
metric
and other products for ordinary sacchariFor saccharimeters employing a normal weight of 26 gms., 0.01 degree Ventzke corresponds to 0.0026 gm. sucrose in 100 true cubic centimeters. Since the majority of saccharimeters can
of sugar, molasses,
work.
be read only to 0.05 V it is evident that weighing within 5 mgs. is sufficiently accurate for ordinary purposes of saccharimetry. The weighing out of normal weights of sugar, etc., for saccharimeters should
not be done upon an analytical balance; the errors due to evaporation from moist substances during the slower adjustment of the analytical balance will usually exceed any advantage in greater accuracy of " " weight. A so-called of the type shown in Fig. 116 sugar balance
163
answers very well for this kind of work. This balance may also be used for the weighing out of chemicals for making up solutions of A set of second quality weights should be provided for apreagents. proximate weighing, and also the normal weights belonging to the
saccharimeter.
The Mohr cubic centimeter normal and half-normal weights (26.048 gms. and 13.024 gms.) are usually furnished in a cylindrical form and the true cubic centimeter weights (26.000 gms. and 13.000 gms.) in a
cubical form (Fig. 123), the shape of the weight thus guarding against
Fig. 117.
Metric solution
scale.
Normal frequently upon the

confusion.
be tested weights, which are in constant use, should
analytical balance against losses in weight through a deficiency exceeding 1 mg. is noted, the stem of the weight should be unscrewed and a small piece of tin or aluminum foil be placed in the cavity sufficient to bring the weight up to its proper value. In addition to the two balances just described a heavy balance or
wear.
If
of weighing out material in bulk, preparing large quantities with sliding A metric solution scale reagents, etc., will be required. is very good for this purpose. 117 in is shown such as Fig. counterpoise A set of third quality weights up to 5 kgs. should also be provided for
scale for
coarse weighings.
FLASKS FOR POLARISCOPIC
WORK
For the preparation of sugar solutions in polarimetric and saccharimetric work various flasks have been devised of different shape
and
construction.
164
SUGAR ANALYSIS
When sugar solutions are made Flasks for Solution by Weight. a to glass-stoppered flask of the form shown percentage up according The flask, which is supplied in recommended. 118 is in VI as No. Fig. Before be not need using, it is thoroughly graduated. many sizes,
cleansed
and
dried,
and then weighed.

is
substance to be examined
The approximate quantity of then transferred to the flask and after
tilled
stoppering the latter is reweighed. The approximate amount of diswater or other solvent is then added and the flask and contents
The percentage of substance in solution is reweighed as before. then readily calculated from the weight of substance taken and the combined weights of substance and solvent. The flask should not be filled too full; sufficient space should be left for gentle rotation of the
liquid
while effecting solution. stoppered to prevent evaporation.
The
flask
should always be kept
Reduction of Solution Weights to Vacua. For very accurate physical measurements the weights taken in air must be reduced to vacuo, since a substance weighed in any medium loses in weight an amount
equal to that of the
medium
displaced.
If
is
the true weight of a

is
substance of density D, in vacuo, then the volume of substance
-~>
and
if
s is
the density of the air at the time of weighing, the loss in

air will
is
weight of the substance in

of the
loss of
be
sW
-
-jr-
Similarly
if
is
the value
weights in vacuo and d
the density of their material then the
the weights in air will be
sP
-7-
The equilibrium upon the pans
of
the balance between substance and weights in air will then be represented by the equation
whence
W=P
i--
as the weight of 1 c.c. of without sensible error. When brass weights are used (d = 8.4), the weights in vacuo of glass, water and sugar are found as follows: for glass (D = 2.5) = 1.000337 P, for water 20 C. (D = 0.998234) = 1.001061 P, for cane = 1.000612 P. The sugar (D = 1.59),
air
The mean value 0.0012 gm. may be taken
following example will illustrate the
method
of application.
Weight Weight
of flask
165
35.2326 gms. 25.1240 gms.
sugar in air
of flask alone in air
Weight of sugar in air Weight of sugar in vacuo = 10.1086 Weight of flask + sugar + water in Weight of flask + sugar in air
X
air
1.000612
10.1086 gms. 10.1148 gms.
95.3055 gms. 35.2326 gms.
Weight Weight Weight
of water in air 20
of water in of sugar
vacuo water
C = 60.0729 X in vacuo =
1.001061
60.0729 gms. 60.1366 gms.

70.2514 gms.
Per cent sugar in solution from weights in air = 14.403 per cent. Per cent sugar in solution from weights in vacuo = 14.398 per cent.
It will
be noted that the difference
is
exceedingly slight, so that
weighing in air is sufficiently exact for all operations except those de-
manding extreme accuracy. Volumetric Sugar Flasks. When solutions of dissolved sugars are made up to a definite volume before polarization, a carefully calibrated volumetric flask must be used; such flasks are supplied in a If solutions are polarized immediately variety of forms and sizes.
making up to volume, as is usually the case, that the flask be fitted with a glass stopper.
after
it is
not essential
'1
III
JV
V
in 10-c.c., 20-c.c., 25-c.c.,
Fig. 118.
Types
of flasks for polariscopic analysis.
Volumetric flasks for sugar work are

50-c.c., 100-c.c., 200-c.c.,
made
and
are also occasionally used. of insoluble matter is allowed for, flasks of irregular capacity are used, as 100.6-c.c., 201.2-c.c., etc., for polarization of sugar-beet pulp.
250-c.c. sizes; 500-c.c. and 1000-c.c. flasks For certain kinds of work, where volume
few of the more ordinary forms of sugar flask are shown in Fig. These may be obtained of any desired capacity. Small sized stoppered flasks similar to No. I are convenient for preparing solutions when small amounts of substance are available. Kohlrausch's sugar
118.
166
flask (No.
SUGAR ANALYSIS
IV) with enlarged top is convenient for transferring substances and is in many ways a most desirable flask; it can be obtained in the small sizes and, if desired, with ground-glass stopper. Sugar flasks with double graduation (No. Ill) for one-tenth dilution are useful for the methods of inversion; they are supplied in 25-27.5-c.c., 5055-c.c., 100-110-c.c.,
and 200-220-c.c.
sizes.
In the selection of sugar flasks Sugar Flasks. the following requirements of the United States Bureau of Standards for volumetric flasks will be found useful. " and of all flasks must be circular
Specifications for
The
cross section
throughout
the neck must merge into the body of the flask so gradually that there Flasks that are manifestly will be no hindrance to uniform drainage. The in construction defective will be rejected. otherwise or fragile
part on which the graduation
mark is placed must be transparent, of The graduation mark must be uniform thickness, and free from striae. the end and not less than 2 cm. less than 6 cm. from not upper placed from the lower end of the neck of a flask larger than 100 c.c., and not
than 3 cm. from the upper end or 1 cm. from the lower end of the neck of a flask not larger than 100 c.c. The graduation mark must extend entirely around the neck. The bottom of the flask must be slightly reentrant, and the flask must be of such form that drainage can take place from the whole interior surface at the same time. The neck of a flask must be perpendicular to a plane tangent to the bottom. The flask must stand solidly when placed on a horizontal plane."
less
A very desirable
in
No.
II, Fig. 118,
Laboratories of
work is that shown Custom-House the United States Treasury Department. The pear100-c.c. flask for saccharimetric
and
in Fig. 123 designed for use in the
shaped body with its low center of gravity gives the flask greater stability than a spherical form. According to the regulations of the Treasury Department "the flasks shall have a height of 130 mm.; the neck shall be 70 mm. in length and have an internal diameter of not less than
mm. The upper end of the neck and the graduation marks shall be not less than 30 mm. from the upper end and 15 mm. from the lower end of the neck." With this size of flask the base of the thumb can cover the mouth and the fingers of the same hand easily enclose the bottom a feature
11.5
mm. and
be
not more than 12.5
shall
flared,
of great convenience
when mixing the contents
after
making up
to
volume.
Calibration of Sugar Flasks. Sugar flasks are graduated to contain 100 true cubic centimeters at 20 C. or 100 Mohr cubic centimeters at 17.5 C. and should be calibrated before using in the follow-
ing manner.
167
The flask to be tested is first thoroughly cleaned and then dried, weighed empty at the temperature of standardization, and then again when filled to the mark with distilled water at the standard temperature. The distilled water should be boiled just before using, in order to expel dissolved air, and then cooled. Special
necessary in adjusting the meniscus to the graduation mark; the lowest point of the curve when
care
is
viewed against a white surface should just touch the graduation mark, the latter appearing to the eye in proper position as a straight line and not as an ellipse. Fig. 119 indicates the proper method of The inside of the neck above the meniscus adjustment. should be wiped perfectly dry with filter paper before reweighing; air bubbles should not be allowed to adhere
level of the
to the walls of the flask during calibration.
Fi g< 119.
Showing
Volumetric 100-c.c. sugar flasks graduated according proper adjustment to the Mohr system should contain 100 gms. of distilled of meniscus,
water at 17.5 C., when weighed in

flasks
air against brass weights; 100-c.c. true cubic centimeters should contain to graduated according 100 gms. of distilled water at 4 C. when weighed in vacuo or 99.7174
vacuo
gms. at 20 C. when weighed in air with brass weights. (Weight in of 100 c.c. water at 20 C. is 99.8234 gms. and weight in air = 99.7174 gms.) The grams of water (p. 164) is 99.8234-^ 1.001061
contained by the flask at 20 C. plus the correction 0.282 will give the volume in true cubic centimeters. The limits of error allowed by the United States Bureau of Standards for volumetric flasks are the following:
Capacity.
168
lot of
SUGAR ANALYSIS
200 sugar flasks purchased by the New York Sugar Trade Laborathe following errors upon calibration. showed tory
Error
in
volume.
not allow sufficient evaporation, during the necessary time of tion, to cause any appreciable error in the polariscope reading.
169
filtra-
MOUNTING OF POLARISCOPES AND CARE OF APPARATUS

If
the circumstances permit, polariscopes should always be
mounted
in a separate room or compartment, where there is no danger of corrosion from the action of fumes or vapors. The polarizing compartment should be well ventilated and easily darkened; lamps and burners
for illumination should be placed
upon the opposite
side of
a wall or
partition.
Fig. 121.
Cabinet for constant temperature polarization (New York Sugar
Trade Laboratory).
In the
(Fig.
New York
Sugar Trade Laboratory the polariscope cabinet
room. 121) constitutes a section of the constant-temperature The roof of the cabinet is composed of shutters, for regulating the downward of cool air, and the sides of the cabinet are enclosed
passage
by dark
leave a space of one foot at the bottom. This arrangement allows free circulation of air, and the presence of several observers in the cabinet does not affect the temcurtains, which,
when drawn,
perature.
170
SUGAR ANALYSIS
is
Where room
polariscopes may laboratory as shown in Fig. 122.
not available for a separate compartment, the be mounted in a large box in a dark corner of the
The
By
table supporting polariscopes should be of solid construction. placing the table upon rubber cushions and setting the polariscopes
of the instruments
upon rubber mats, vibration
and consequent
dis-
turbance of the zero point will be largely obviated.
Fig. 122.
Portable polariscope cabinet with section of side removed.
It is essential in saccharimetric
work that
all
apparatus be kept
scrupulously clean. The more delicate optical parts of polariscopes, such as polarizer, analyzer, and quartz compensation, are enclosed, in the most modern apparatus, in dust-proof housings, and very rarely The diaphragm glasses (A and P, Fig. 96) at require to be disturbed.
each end of the polariscope trough are the parts which require most
attention. Drops of solution, accidentally adhering to the polariscope The tubes, are occasionally splashed against the diaphragm glasses.
171
diaphragms, which either screw or slide into position, should be examined frequently and the glasses wiped free of dirt and dust particles. A paper napkin will be found very suitable for cleaning diaphragm
glasses,
The troughs
quently become
eye pieces, and other exposed optical parts. of polariscopes in the hasty round of routine
soiled
fre-
from contact with wet tubes or spilled liquid. be should wiped frequently with a damp cloth and the metal They surface should be kept smooth and clean. The bichromate cell should be examined frequently, and the solution replenished as soon as bubbles begin to form, otherwise their
appearance
may
obscure the
field.
the polariscope is not in use, the trough should be closed and the instrument kept covered.
When
must also be observed in the use of polariscope and other In handling and carrying obseraccessories. tubes, flasks, vation tubes a portable rack of the form shown in Fig. 122 will be
Strict cleanliness
found convenient.
Where sugar solutions are clarified with lead subacetate, the walls of flasks, cylinders, funnels, and tubes become coated in time with a thin white film of lead carbonate. good solvent for this coating is a
warm
solution of
sodium hydroxide and Rochelle
salts,
such as
is
used
in preparing Fehling's solution. Hydrochloric or nitric acid may also be used for removing the deposit. After thorough rinsing in clean
water, tubes, flasks, funnels, and dry upon racks.
and
cylinders should be allowed to drain
CHAPTER
VIII
struction
IN the previous chapters the principles which underlie the conand operation of polariscopes were described; it is now de-
sired to study the application of these principles to of sugar analysis.
some
of the
problems
The
specific
power of a sugar is expressed as specific rotation, or rotatory power, by which is meant the angular rotation which
polarizing
a calculated 100-per cent solution, 1-dcm. long, gives to the plane of

polarized light. The specific rotation, indicated by the expression [a] can easily be calculated from the angular rotation a of the solution of
substance by means of the equation
[a]
=
c
= > l
in
which
c is the
con-
centration of substance (grams per 100-c.c. solution) and I the length Instead of the foregoing we of the observation tube in decimeters.
may
use the equation
[u]
=
J)
^ X X CL
>
in
which p
is
the percentage of
substance in solution (parts by weight hi 100 parts by weight of solu= c in previous tion) and d is the specific gravity of solution, (p X d
equation.)
rotation, as the of length light employed.
The angular
used for polariscopic
shown below, depends upon the wave Sodium light is the illumination most measurements and as the bright yellow line of
sodium is designated the D line of the solar spectrum, the expression [a] for sodium light is written [a]b. Specific rotation for the mean yellow no The temperature at which is written (now ray j longer used) [a]j. the specific rotation is taken is also usually affixed. Thus: the symbol for specific rotation using sodium light at 20 C. is written [a]. The method of calculating specific rotation may best be understood by an example; 20 gms: of cane sugar dissolved to 100 c.c. gives an angular rotation for sodium light of +53.2 degrees in a 400-mm. tube at 20 C.
Substituting
these
values
in
the
equation
[a]
=
c
=->
we obtain
given
MD =
100 X 53 2 on x 4
= ~^~ ^'** *ê
specific rotation of sucrose for the
concentration.
172
173
To
calculate specific rotation
from the reading
the scale divisions of the latter
must
first
degrees by means of the appropriate factor. dissolved to 100 metric cubic centimeters gave a reading of +57.7 in a 200-mm. tube using a Ventzke scale quartz-wedge saccharimeter.
Since 1
of a saccharimeter, be converted to angular Thus: 15 gms. of sucrose
V=
r
0.34657 angular degrees (page 145) then

,
[a] D
sucrose
100 (0.34657 X 57.7) 15 x 2
+ 66.6.
EFFECT OF KIND OF LIGHT UPON SPECIFIC ROTATION OF SUGARS.

Mention has been made
specific rotation.
of the influence of
wave length of
light
upon
In Table
XX a comparison was given of the rotations
of quartz
and sucrose for light of different wave lengths and it was shown that as the wave length decreases the polarizing power of sucrose
In the following table the specific rotations of nine different sugars are given for light of different wave lengths in the red, yellow, green, blue, indigo, and violet parts of the spectrum, according to
recent measurements
increases.
by Grossmann and Bloch.*
The
specific rota-
tions for yellow sodium light, [O\D, the standard values of comparison, are printed in heavier type.
Sugar.
174
SUGAR ANALYSIS
The
results as
expressed in ten-thousandths of a millimeter
thus calculated have only an approximate value, as other factors, such as temperature, concentration, etc., are not considered.
The
specific
rotations of the different sugars also vary according
to the concentration of solution, the temperature of observation
and
the nature of the solvent.
following table gives the approximate values for the specific rotation of a number of sugars. The effect of concentration and temperature in increasing or lowering the specific
rotation
is
The
indicated
by the
direction of the arrow in the respective
columns.
TABLE XXVII
Showing Effect of Increase in Concentration and Temperature upon Specific Rotation
of Sugars
Sugar.

I.
175
II.
For the straight line For the parabola For the hyperbola
[a]
[a]
= =
III.
p determined the and nature of the curve it remains Having plotted to calculate the values of the constants a, b, and c in the above equations. The method of doing this (the method of least squares) is simple, although the work of calculation is somewhat laborious. The following example is given as an illustration
:
[a]
+ bp. + bp + cp = a + -^ C +
a
a
2
.
From
Nasini,
and Villavecchia, the following

40 per cent
the average results of observations by Tollens, Thomson, Schmitz, specific rotations of sucrose were found
for different concentrations; 10 per cent
cent
+ 66.41,
+ 66.56, 20 per cent + 66.52, 30 per + 66.27, 50 per cent + 66.06. An equation desired
is
for the specific rotation of sucrose for
any concentration within these
limits.
By plotting the above observations a curved line is obtained presumably a parabola. (In calculating the concentration curves for the specific rotation of sugars the hyperbola is but little used.) Substituting the results in the previous
equation II for the parabola we obtain the following
1.
:
2.
3. 4.
5.
Average:
I.
+ 10 b + a + 20 b + a + 30 b + a + 40 b + a + 50 b + a + 30 6 +
a
100
400 900 1600 2500 1100
c c c
c
= = = = = =
66.56.
66.52. 66.41. 66.27. 66.06.
66.364.
From
the above equations

6. 7.
we obtain by
40 b 30 b 20 6
30 b
subtraction the following:

0.50. 0.46. 0.35.
8. 9.
10.
11. 12.
13. 14. 15.
(5-1) (5-2) (5-3) (5-4) (4-1) (4-2) (4-3)

(3
+ 2400 c = + 2100 c = + 1600 c = 900

1500 1200
c
106+
20 6
=-0.21.
206+ (3-2) 106+ - 1) 10 6 + (2

1)
+ + 10 b +
c c
c c c
700
===-
0.29. 0.25. 0.14. 0.15. 0.11. 0.04.
800 500 300 1200
====-
c c
Average:
II.
20 6
0.25.
By combining
the following:
III.
equations 6 to 15 into two series and subtracting
we
obtain
(7
IV.
+ 8 + 10 + 12 + 14) (6 + 9 + 11 + 13 + 15)
100 6
100 6
+ 6400 c = + 5600 c =800

c c
1.35
1.15
==-
0.20
0.00025.
176
SUGAR ANALYSIS
= 0.0025, and Substituting the value for c in equation II we obtain b = SubI a 66.564. we obtain c in b for and values these equation substituting obtain: we for the in the values these parabola original equation stituting
[a]g
66.564
0.0025 p
0.00025 p\
The calculated specific rotation of sucrose for various concentrations according to the above equation is as follows: 10 per cent 66.56, 20 per cent 66.51, 30 per
cent 66.41, 40 per cent 66.26, 50 per cent 66.06, results which agree perfectly with the average observations taken.
The above equation for the specific rotation of sucrose does not hold, however, for concentrations below 10 per cent or above 50 per cent. Tollens * from observations upon 19 solutions ranging from 3.8202 per cent to 69.2144 per cent sucrose calculated the following equations:
For p
= =
For p
4 to 18 per cent sucrose, - 0.015553 p [] = 66.810 18 to 69 per cent sucrose,

2
0.000052462 p 2
H D = 66.386 + 0.015035 p - 0.0003986 p\

=
According to the above equations the maximum specific rotation of sucrose (66.53) is found at p = 18.86 per cent; for concentrations
lower than this the specific rotation again decreases. Schmitz | from observations upon eight solutions for p per cent gives the equation:
[a]
5 to 65
66.510
+ 0.004508 p |
0.00028052 p\
to
.
Nasini
[a]
and Villavecchia
for
p =
65
give
last
(c
the
66.438
0.010312 p
0.00035449 p 2
The
named
equation au-
thorities found,
however, for very dilute solutions
0.335 gm. to
1.2588 gms. sucrose per 100
again increases, and for = 69.962 - 4.86958 p []g
c.c.) that the specific rotation of sucrose such dilute solutions give the equation 1.86415 p 2 The variations noted in the
.
above equations for the specific rotation of sucrose are no doubt partly due to the effect of rotation dispersion, as the result of using light of
wave length for illumination. The equations of Tollens and of Nasini and Villavecchia are considered to be the most accurate. The average of the two equations gives
slightly different
probably the most reliable expression for the specific rotation of sucrose. 2 = I66.386 0.015035 p - 0.0003986 p 2 [a] D (Tollens.)
II.
[a]
= +66.438 + 0.010312 p - 0.0003545
Average:
III.
*
[]
= +66.412 +
p (Nasini and Villavecchia.) 0.012673 p- 0.0003766 p 2

.
Ber., 10, 1403. t Public, de lab. chim. delle gabelle.
Ber., 10, 1414.
Rome,
1891, p. 47.

Landolt
*
177
of
by
recalculating this
combined equation into terms

c.c.)
concentration (grams of sugar per 100

IV.
gives the expression:

c (c
2
[]
= + 66.435 + 0.00870 c - 0.000235
to 65).
The following table, which with the exception of column / is taken from Landolt,* gives a comparison of the specific rotation of sucrose
for solutions of different percentage and concentration, according to each of the four preceding equations.
TABLE XXVIII
Giving Specific Rotation of Sucrose for Different Concentrations
a
178
SUGAR ANALYSIS
EFFECT OF TEMPERATURE UPON SPECIFIC ROTATION OF SUGARS
The
no
less
temperature upon the specific rotation of sugars is pronounced than that of concentration and, with a number of
effect of
sugars such as fructose and galactose, the influence of temperature is the factor which has most to be considered in polarimetric measurements. The change in rotation of a sugar solution due to expansion
or concentration in
volume through temperature changes must not be
confused with changes in specific rotation. In studying the latter phenomenon the sugar solutions must either be made up to volume at
the same temperature at which they are to be examined or else a correction be made for the changes in volume due to expansion or contraction.
The influence of temperature upon specific rotation is studied in the same way as that of concentration, by laying off the specific rotaThe connecting points for tion for each temperature upon a diagram. of the ordinary ranges atmospheric temperature lie more nearly in a
straight line than
is
the case with the concentration curves.
For
wider ranges of temperature, however, the increase or decrease in specific rotation is found to proceed unequally and the change must
then be expressed by some curve equation. Effect of Temperature upon the Specific Rotation of Sucrose.
Hesse, and Tuchschmid reof effect the the temperature upon specific rotation of sucrose garded Dubrunfaut* was the first to recognize the fact as insignificant.
The
earlier investigators Mitscherlich,
that increase of temperature caused a decrease in the value of this constant, the temperature coefficient of the specific rotation of sucrose having been
Andrews,! who
found by him to be 0.000232 per 1C. increase. reinvestigated the question in 1889, found a decrease
1
t
of 0.0114 in the specific rotation of sucrose for specific rotation of sucrose for any temperature
C. increase.
is
The
then represented
by the equation:
MA = MS -0.01140
tions,
-20).
SchonrockJ upon 10 sugar solushowed that the decrease in specific rotation for 1 C. increase lay between 0.0132 and 0.0151 for temperatures between 12 C. and 25 C. the change is expressed by the equation:
;
in 1896, as a result of observation
MA = MS ~
*
0144
(*
20).
Ann. chim. phys. [3], 18, 201. Mass. Inst. Tech. Quarterly, May, 1889,
phys.-techn. Reichsanstalt, 1896.
p. 367.
t Ber.

This equation
is
179
sometimes written
Mb =
in
[<*}D
~ MS 0.000217
(t
20),
which the temperature
coefficient of the specific rotation,
000217
--
0144
66.5
Later experiments were made by Schonrock* at temperatures between 9 C. and 32 C. using light of three different wave lengths, the yellow sodium line 589.3 w, the yellow-green mercury line 546.1 "/*/*, and the blue mercury line 435.9 /*/*. These experiments showed that for the German normal sugar solution (p = 23.701 per cent) the rotation angle underwent a linear deviation with changes in temperature, this
deviation being independent of the wave length of light employed. It was found, moreover, that the temperature coefficient of the specific
rotation decreased with increase in temperature, the value being 0.000242 at 10 C., 0.000184 at 20 C., and 0.000121 at 30 C. for Sodium light.
This decrease proceeds in a straight line and the values of the temperature coefficient for any intermediate temperature can be estimated by
taking the proportionate difference. These later values of Schonrock are used by the Physikalisch-Technische Reichsanstalt of Germany and have therefore the highest sanction of authority.
-
of
Temperature upon the Specific Rotation of Other Sugars. temperature upon the specific rotations of a number other sugars is given in Table XXIX.
Effect of
The
effect of
TABLE
XXIX
9.18-0.035
84.67-0.200
t
Rhamnosef
Fructose
................
GalactoseJ (p = 10) .......... []
+ H= =
(1= 6
(<
(/.
to 20 to 30
C.)
(p=9) ............
H^ = - 103. 92+0. 671
= 10 = 13
C.)
C.)
to 40 to 45
to 35
Fructose
(p=23.5) ......... [a]^ =

(c
Invert sugar||
= 17.21) .....
+ 52.53-0.07 i-20 (* = 15 to 25 C.) Lactose^ ................... [! Maltose** (p=10) ........... [J^ = + 140. 19 -0.095 t (* = 15 to 35 C.)
* t
W=
[2],
- 107. 65+0. 692 = 27.9 +0.32 D
(1= 9
(t= 5
C.) C.)
Z. Ver. Deut. Zuckerind., 53, 650. Schnelle and Tollens, Ann., 271, 62.
j Meissl, J. prakt.
Chem.
22, 97.
2, 235.
Honig and
||
Jesser, Z. Ver.
J.
Deut. Zuckerind., 38 (1888), 1028.

[2],
Tuchschmid,
prakt.
Chem.
[2],
i Schmoger,
**
Ber., 13, 1922.
Meissl. J. prakt.
Chem.
25, 114.
180
SUGAR ANALYSIS
is
While a linear equation
sufficiently exact for
narrow ranges
of
tem-
of temperperature, the change in specific rotation for wider differences ature must usually be expressed by an equation of the order:
or
[<x}'
[<*}%
+ a(t-20)+b(t- 20)
0.03642
t
2
.
Gernez,* for example, gives for rhamnose the equation

\aY D
9.22
+ 0.0000123
and Gubbef gives

For
*
for invert sugar the following equations:

.
For
= = 20
to
30 C., [a]^ = [ag+ 0.3041 (t - 20)+0.00165 (*-20) 2 2 to 100 C., [*] = []%+ 0.3246 0-20) -0.00021 0-20)
Sucrose and the different sugars mentioned in Table XXIX all show a decrease in specific rotation with increase in temperature. Of other sugars, which exhibit this property in marked degree, arabinose should
be mentioned.
TanretJ found
for 1-arabinose [a]^
= +105.54 and
[a]g= +88.61, or an average decrease of 0.394 for 1 C. increase in temperature, which is greater than that for any other sugar except
fructose.
Xylose presents an exception to the rule just noted, Schulze and having observed for temperatures above 20 C. an increase in = 10.0829). specific rotation, as in the following example (p
Tollens
t

Galactose * [] Fructose [alp
Fructose
181
= + 83.883 + 0.0785 p - 0.209

= = [101.38
1.
(Meissl.)t
10)].
0.56
0.
108
(c
(Jungfleisch
and
Grimbert.)t
[]
88.13
0.2583 p
+ 0.6714
(t
20).
(Honig
and
Jesser.)
Sorbose
[]g =
[42.65
+ 0.047 p + 0.00007 p - (t- 20) 0.02].

2
II
(Tollens
and Smith.)
Maltose
[]g =
+ 140.375 - 0.01837p - 0.095
1.
(Meissl.)
1F
EFFECT OF SOLVENT UPON THE SPECIFIC ROTATION OF SUGARS
The constants
aqueous
solutions.
of specific rotation for sugars are all expressed for It sometimes happens, however, that solutions of
sugar in other solvents, such as alcohol, have to be examined; in such cases the changes in specific rotation due to the character of solvent
must be taken
into account.
In the case of sucrose, Tollens** found the following values for [a]^ with different solvents for a 10 per cent solution:
In water
In In In
1
1 1
+ 66.667. + 3 parts ethyl alcohol + 66.827. part water + 3 parts methyl alcohol + 68.628. + 67.396. part water + 3 parts acetone
part water
raise the specific rotation
Methyl alcohol and acetone are thus seen to

of sucrose perceptibly,
but ethyl alcohol only slightly. Claassenft also found for 80 per cent alcohol a slight increase in the specific rotation of sucrose; the differences (0.1 to 0.15), however, are not sufficient to affect seriously the analytical results in such operations as the alcoholic extraction of sugar beet or cane pulp. In the case of fructose and invert sugar, ethyl alcohol produces a
of the specific rotation, and when these sugars are the influence of ethyl alcohol as a solvent must be taken into present
marked lowering
*
rotation of galactose for
change in specific -0.39, between 20 and 25 -0.226, and between 25 and 30 -0.180, a falling off in the temperature coefficient with increase in temperature similar to the one noted by Schonrock with
(Bull,
societe"
1
Tanret
chimique
[3],
16,
195) gives the
C. increase between 13
and 20
sucrose.
t J. prakt. t
Chem.
[2],
22, 97.
Compt.
Z. Ver.
rend., 107, 390.
Deut. Zuckerind., 38 (1888), 1028.
II
Ber., 33, 1289.

J.
1f
prakt.
Chem.
[2],
25, 114.
**
Ber., 13, 2287.
ft Z. Ver, Deut. Zuckerind., 40, 392.
182
account.
SUGAR ANALYSIS
Fructose according to Landolt* has a specific rotation in is only two-thirds that in water. Borntrsegert found for 49.2 37.6 gms. invert sugar in 100 c.c. aqueous solution a rotation of at 20 C.; when the solution was made up with 10.45 c.c. alcohol 38.3. 43.9 and with 20.60 c.c. alcohol to the rotation decreased to
alcohol which
According to Horsin-Deon t (whose conclusion, however, requires verification) invert sugar in absolute alcohol is perfectly inactive and only becomes levorotatory upon the addition of water. It should also be
noted that the rotation of alcoholic invert-sugar solutions is more sensitive to changes in temperature than water solutions.
much
of sugars the specific rotations in aqueous and The [O\D alcoholic solutions are almost the reverse of one another. in is in water and alcohol 9.0. for example of rhamnose +9-43
With a number
The
[O\D of sorbosej
in
+41.8.
The
effect of pyridine
is
tions of several sugars
42.5 and in 85 per cent alcohol is and formic acid upon the specific rotashown on page 190.
water
Without giving detailed

it
results of experiments
effect of solvent
may sugars is too great to be disregarded;

solution.
be said that the
upon all the various upon specific rotation
wherever possible the polarimetric ex-
amination of sugars for purpose of analysis should be
made
in
aqueous
EFFECT OF ACCOMPANYING SUBSTANCES UPON SPECIFIC ROTATION OF SUGARS

Another factor of importance, especially in the polarimetric examination of impure sugar solutions, is the effect which bases, acids, salts, and other substances exert upon the specific rotation of the sugars
present.
this subject
and
very large amount of investigation has been done upon for complete details reference must be made to the
original articles. Only brief mention will be made of the effects of a few substances upon the rotation of the more important sugars. The changes which foreign optically inactive substances may exert upon the rotation of sugars may be either chemical or physical. The
hydroxides of the alkalies and alkaline earths, and all salts of alkaline reaction in general, cause a decrease in the specific rotation of most
reducing sugars.
*
Such changes in rotation are purely chemical, being
Ber., 13, 2335. t Z. ang. Chem. (1889), 507. J J. fabr. sucre, 20, 37.
II
Rayman and Kruis, Bull. soc. chim. [2], 48, 632. Adrian!, Rec. trav. chim. des Pays Bas., 19, 184.

due
183
either to a rearrangement of the sugar molecule or to the forma-
tion of alkali-sugar compounds of lower specific rotation. The effect of acids and acid salts upon the rotation of sucrose by inversion is another
example of purely chemical change. The avoidance of such chemical changes is imperative in accurate polarimetric work and to prevent these the solutions of sugar under examination should be, so far as
possible, neutral in reaction.
The influence of neutral salts upon the specific rotation of sugars, on the other hand, is largely physical, since the chemical properties of the dissolved sugars are not appreciably affected; the same is also true of
the influence of acids
upon the
specific rotation of sugars
which do not
undergo inversion.
Influence of Mineral Impurities upon the Rotation of Sucrose.
The
chlorides, nitrates, sulphates, phosphates, acetates, and citrates, of the alkalies, the chlorides of the alkaline earths, magnesium sulphate,
and many other
salts all
sucrose, this decrease being generally greater
produce a decrease in the specific rotation of with increased amount

of salt.
and smaller molecular weight
The hydroxides
of the alkalies
and
alkaline earths
and the carbon-
ates of the alkalies also lower the specific rotation of sucrose. The influence of these substances, which is of especial importance technically,
in
view of the alkalinity of various sugar-house products, has been
widely studied, the results being often expressed in parts of sugar whose rotation is obscured by one part of alkali. Pellet for example gives
the following results:
Substance.
184
SUGAR ANALYSIS
a
slight extent.
however, since the soluble acetates themselves lower the specific rotation of sucrose to
The probable
sucrose,
effect of
a mixture of
salts
upon the polarization
of
such for example as occurs in beet molasses, which contains about 50 per cent of sucrose and 10 per cent of soluble salts (mostly of may be judged from the following examples taken from potassium),
experiments by Bodenbender and Steffens.*
TABLE
Salt.
XXX
185
metric analysis the influence of lead subacetate, as a clarifying agent, upon the rotations of fructose and invert sugar, is f great importance.
first observed by Gill* in 1871 when solutions containing invert treated with lead-subacetate solution in excess, the formation are sugar of soluble lead fructosate of low specific rotation is so pronounced that
As was
the rotatory power of fructose sinks below that of glucose and the invert sugar becomes dextrorotatory. Similar observations have been
made by
In Pellet, Bittmann, Koydl, Borntraeger, and many others. the following experiments by Bittmann f 50 c.c. of invert-sugar solution were treated with 50 c.c. of a mixture of water and lead subacetate in
different proportions.
Water.
186
SUGAR ANALYSIS
has calculated to the [a]g of invert sugar and fructose. The results Were obtained by inverting a half-normal weight of sucrose with varythen completing ing amounts of concentrated hydrochloric acid and
the volume to 100
c.c.
TABLE
XXXI
the Rotation
Showing Influence of Varying Quantities of Hydrochloric Acid upon of Invert Sugar and Fructose.
Volume
of
HC1 added.
187
arabinose, arabinose and xylose also show that it is safe to assume in analytical work that the specific rotation of these sugars is not per-
ceptibly affected
by other sugars
in solution.
MUTAROTATION
observed in the polarization of all optically active that of mutarotation (also called birotation or multireducing sugars The rotation). polarizing power of such sugars undergoes after soluis
A phenomenon
tion at first a rapid change which slowly

after
becomes more gradual until a few hours the polariscope reading remains constant. This phenomenon was first observed upon glucose in 1846, by Dubrunfaut * and the fact that the initial rotation of this sugar was about twice the constant value caused the introduction of the name birotation. The relation 2
:
was found, however,
to be different in the case of other
sugars; Wheeler and Tollens,f for example, found the ratio in case of xylose to be about 4.5:1 and accordingly suggested the name multirotation.
This term, however, in recent years has given place to the
more expressive word mutarotation (Latin mutare = to change) introduced by Lowry 1 in 1899. The effect of mutarotation upon the rotatory power of sugars is shown in the following table, in which results are quoted from the work of Tollens and his coworkers, giving the specific rotation of a number of sugars directly after solution and after standing until no further change was noted. The time after solution is given after each value
for
[].
TABLE XXXII
Showing Mutarotation of Different Sugars
Sugar.
188
It is
SUGAR ANALYSIS
noted that in case of rhamnose there is a decrease in rotation to 9.4. Maltose also and then an increase from 5.0 to time of solurotation at less a from the other sugars in showing
from
differs
tion than after standing. Effect of Temperature
on Mutarotation.
The speed
of
muta-
rotation
by
is influenced by a large increase in temperature, the change proceeding very slowly at Dilute sugar solutions show the C., and almost instantly at 100 C.
number
of factors.
It is accelerated
change for all concentrations. Highly concentrated solutions, however, do not always give the true end rotation; such solutions must first be diluted and then allowed to stand for the change This fact must be borne in mind in the in rotation to be completed.
same velocity
of
polariscopic examination of concentrated sugar solutions, such, for example, as liquid honey, otherwise a considerable error may be intro-
duced in the work of analysis. Velocity of Mutarotation.

initial to
The
velocity
of
the change
from
and also Urech * varies according to temperature, solvent, and other conditions. was the first to show that the speed of mutarotation followed the same
constant rotation
is
different for different sugars,
law as that noted by Wilhelmy in the inversion of sucrose (page 660), and which is expressed by the following general formula for a reaction
of the first order,
-
k (a
- x),
in
which k
is
beginning and end
the coefficient of velocity, a the total change between the point, and x the change at the end of any time t.
integration gives
The above equation by
1, t
log &
to the impossibility of measuring the specific rotation of a sugar at the exact moment of solution, the velocity of mutarotation is
Owing
generally determined
by the modified formula
in
which
ft
and
ft are
<f>
times h and ^, and
the rotations at the end of the corresponding the constant end rotation.
is
The method
which
is
of calculation
shown by the following example,
taken from the work of Levy,f

* Ber., 16, 2270; 17, 1547; 18, 3059. t Z. physik. Chem., 17, 301.

TABLE XXXIII
Showing Velocity of Mutarotation for a Glucose Solution = 1.0114. Temperature.= 20.5 to 20.9 C. Per cent, C6 H 12 O 6 = 3.502. d
Time
after solution.
189
190
SUGAR ANALYSIS
for relative acceleration of the different acids preserve
The values
the same
(page 663).
order as those noted for the inversion constants in Table
XCV
It is scarcely necessary to state that the speed of mutarotation Thus Levy found for increases with the strength of acid employed.
= 0.02300 and for n/50 hydrochloric acid, ft/10 hydrochloric acid, k k = 0.00971; for n/10 acetic acid, k 0.00716 and for n/50 acetic
acid, k
0.00654.
Alkalies also accelerate the speed of mutarotation, the change to constant rotation being almost instantaneous. Schulze and Tollens* using 0.1 per cent ammonia obtained the normal constant rotation
with arabinose, xylose, rhamnose, galactose, glucose, fructose, and lactose within 9 minutes; n/200 alkali (KOH) gives the end rotation of
The use of much stronger alkali, however, glucose almost instantly. induces chemical change with a decrease of the rotation below the normal
Treyf for example using 0.2 gm. sodium hydroxide per 100 c.c. obtained as the [O\D for glucose after 15 minutes 52.7 (normal), after 24 hours 36.7, after 48 hours 26.0, after 34 days 15.1, and after
value.
65 days
0.4.
The different salts nearly all accelerate the speed of mutarotation, those of alkaline reaction standing first in this respect. Sodium chloride,
by Levy I and
however, presents an exception to this rule, having been found also by Trey to cause the mutarotation of glucose to
proceed slower than in pure aqueous solution. Mutarotation of sugars takes place not only in water but also in o^her solvents such as absolute methyl alcohol, ethyl alcohol, acetone,
etc.
The change
in rotation proceeds
much more
This
is
organic solvents than in aqueous solution.

||
shown
slowly, however, in in the follow-
ing results by Grossmann and Bloch which give the mutarotation of several sugars in pyridine and formic acid.
Sugar.
191
A peculiarity of xylose and rhamnose in pyridine is an increase in the rotation after solution. Grossmann and Bloch observed a maximum 122.07 in case of xylose 15 minutes after solution and a maximum of
45.92 in case of rhamnose 30 minutes after solution. It is seen that mutarotation in the two solvents proceeds in many cases in opposite directions and that there is no relation between the constant rotations
of
and those observed
in
aqueous solution.
The
addition of water to
solutions of sugar in organic solvents accelerates, and conversely the addition of alcohol, acetone, etc., to aqueous solutions retards, the speed
of mutarotation.
As a general
rule the presence of
any
soluble non-
electrolyte, such, for example, as sucrose, will increase the

for a
time necessary
mutarotating sugar to reach constant polarization. Mutarotation takes place not only after dissolving reducing sugars, but also occurs upon the liberation of these sugars from higher saccharides by the action of enzymes. The phenomenon is one which the sugar chemist has always to bear in mind. Polariscopic measurements are always referred to the normal constant rotation. The latter condition may be produced almost instantly by heating the solution or by adding a little free alkali, but when such means are employed care must be
taken to prevent the

is
liability of
chemical change.
to allow the solution to stand until the rotation has
The safest course come to equili-
brium in the natural way. Theories of Mutarotation.
Many
theories have been proposed
to explain mutarotation. According to the views of Landolt* and other authorities it was thought that the phenomenon might be due to the
formation of molecular aggregates immediately after solution, which afterwards decompose into simple molecules of lower rotation. These earlier theories were largely disproved, however, by the experiments of
Arrhenius,t and of Brown and Morris,! who showed that no change occurred in the molecular weight of a sugar during mutarotation. Tollens and others of his school have supposed that mutarotation might be caused by the formation of unstable hydrates which, by the
splitting off of water, cause a
change in rotation.
additional light was thrown upon the subject in 1895 by Tanret, who discovered that sugars could exist in both a high- and a The relationship of these several modificalow-mutarotating form.
||
Much
tions,
according to Tanret's classification, sugars in the following table.

*
is
shown
for four different
optische Drehungsvermogen t Z. physik. Chem., 2, 500. Ber., 26, 1799.
"Das
"
(1879), 58.
t
Chem. News, 67, 196. Compt. rend., 120, 1060.
192
SUGAR ANALYSIS
Sugar.
193
Lowry's view was supported by Hudson,* who showed by quanchange between the high- and low-rotating forms of lactose was a balanced reaction. According to this view,
titative experiments that the
Tanret's solid (3 sugars of constant rotation are simply equilibrated mixtures of the high- and low-rotating forms. The designation /3 is applied at present to Tanret's y modification.
While mutarotation is most generally regarded at present as a balanced reaction between high- and low-rotating forms, the intermediate The change steps of the process have not been definitely established.
in polarization of
a sugar solution to constant rotation is regarded by some chemists as simply a conversion of the a or /3 oxygen ring compound into the ordinary aldehyde or ketone form. Other chemists
regard the solution at constant rotation as containing simply a mixture of the a and /3 sugars in equilibrium, while still others believe it to
contain the a and # sugars with variable amounts of the aldehyde or ketone form. For a review of the different hypotheses, which have
been proposed in this connection, the chemist

special works, f
*
is
referred to the various
Z. physik.
Chem., 44, 487. See also page 711. Lippmann, "Chemie der Zuckerarten" (1904), 293.
Hudson
eries
(J. Am. Chem. Soc., 32, 889) in a paper entitled "A Review of Discovon the Mutarotation of Sugars," gives a very complete review and bibliography
of the subject.
CHAPTER IX
DETERMINATION OF SUGARS FROM ANGULAR ROTATION
a single optically active sugar, in presence of optically inactive substances or in presence of substances without effect upon its specific rotation, may be calculated by means of either formula
of
for specific rotation (page 172).
THE amount
100 a
whence
100 a
100 a
100 a
lXdX[a] D
As
first
to which of the above
methods
is
of calculation is to be used, the
the better where a definite weight of substance is made up to volume before polarization, the usual method of procedure; in case, however, a sugar solution of known specific
or concentration formula
gravity is polarized directly, then the second or percentage formula is to be employed. The following formulae are given for calculating the concentration
(grams per 100

in a 2-dm. tube.
c.c.) of different
sugars from the angular rotation
(a)
Arabinose
4785 a
3.
Glucose
= = =
&
=
10
0.9470
a.
4.
Fructose
X
X
= =
0.5405 a
(left
degrees).
5.
Galactose
0.6173
a.
6.
Sucrose
~
-f-
ol.U
0.7519
a.
194

8.
195
Lactose
=
5
=0.9524 a.
9.
Raffinose
(+
H 0) c =
2
2
c
=
5
0.4785
a.
10.
Raffinose (anhydride)
=
z
T~ iZo.io
is
J*
0.4060
a.
The percentage p
of a sugar in solution
equal to the value of
c,
as expressed above, divided by the specific gravity of the solution. Such formulae, as the above, are sufficiently accurate for most purIn cases, however, where the specific rotation of the poses of analysis.
sugar
is
affected
by changes
in concentration or temperature, the results
as obtained above can be considered only approximate; to obtain the correct concentration or percentage, it is necessary to calculate the
specific rotation corresponding to the approximate value of c or p at the temperature of polarization and substitute this corrected specific rotation in formulae (1) or (2) for the final calculation of c or p.
Example.
50 gms. of a dextrose sirup were dissolved to 100
cc.;
the
constant rotation of the solution thus obtained was -f 34.55 circular degrees in the 200-mm. tube. Required the percentage of dextrose in the sirup.
From formula
we obtain by
substitution c
0.9470
34.55
32.72 gms.
dextrose in the 100 cc. of solution or for the 50 gms. of sirup, 65.44 per cent The specific rotation of dextrose for c = 32.72 is found from approximately.
the formula
[]g
+ 52.50 + 0.0227 c + 0.00022

formula for
5
c <
c2 (p.
177) to be
+53.48;
substituting this in the general
we obtain
=32.30
gm ,
in the
100
cc. of solution or for
0.84 per cent less than the value
the 50 gms. of sirup the true percentage 64.60, by the uncorrected formula.
for the variations c, so as to correct the labor of the second calculation in the above example may be eliminated. In the case of glucose, by calculating the angular rotation, (a) for the 2-dm. tube, corresponding to concentra-
By
modifying the formula for
in specific rotation,
tions ranging
squares
(p.
from 10 to 60, we obtain, using the method of = 0.958 a - 0.00067 a2 165), the formula c*
.
least
Example.
for
c,
Applying the
last
32.299 gms. dextrose in the 100
formula to the previous example, we obtain cc. of solution or for the 50 gms. sirup
64.60 per cent.
For p Landolt gives the formula p hungsvermogen," p. 447.)
0.948 a
0.0032 a 2
("
Optisches Dre-
196
SUGAR ANALYSIS
DETERMINATION OF SUGARS FROM SACCHARIMETER READINGS
Conversion of Saccharimeter Readings into Angular Rotation.
The general methods
pecially applicable to polarimeters,

eters in
of optical analysis just described are more eswhere readings are taken in angular
degrees; the formulae given are equally applicable, however, to saccharimwhich case the scale reading of the latter must be converted
into angular degrees
by means
of the proper conversion factor.
For
general purposes the factor established for sucrose may be applied to In the case of the Ventzke scale, sugar degrees other sugars.
angular rotation. Since, however, the rotation dispersion of the various sugars, with reference to the quartz compensation of the saccharimeter, may differ somewhat from that of sucrose, it is
0.34657
always better, where exact data are available (which is unfortunately not always the case), to use the conversion factor established for the In the case of a few sugars Landolt * has established particular sugar.
the following factors for converting divisions of the Ventzke scale into
circular degrees.
Sucrose Lactose Glucose Invert sugar
Raffinose
0.3465 3452 0.3448 3432 0.3450

. .
Brown, Morris, and Millar f give the following:

Sucrose, 10 per cent solution Maltose, 10 per cent solution Maltose, 5 per cent solution Glucose, 10 per cent solution Glucose, 5 per cent solution Starch products, 10 per cent solution Starch products, 5 per cent solution
'
0.3469 3449 3457 0.3442 3454 0.3458 0.3454

.
. .
Herzfeld,J with a solution containing 11.29 per cent anhydrous maltose, obtained upon a Peters saccharimeter, using a Welsbach light
with chromate filter, a reading of 93.88 Ventzke degrees at 20 C., and with the same solution upon a Lippich polarimeter a reading of 32.60 circular degrees at 20 C. The value of a Ventzke-scale division for
maltose under these conditions
is
therefore
H^ =
9o.oo
0.3471
circular
sugar solutions examined but more especially differences in the optical center of gravity of the light employed for illuminating the saccharimeter are the chief
*
Ber., 21, 194.
f J-
degree, a figure perceptibly greater than the values of and Millar. Differences in concentration of the
Brown, Morris,
Chem.
Soc. Trans., 71, 92.
J Ber., 28, 441.
197
causes of such discrepancies. The chemist should, therefore, employ any prescribed conversion factor with caution and use it only under the
It is also well to verify for which it was established. a conversion factor wherever possible, by comparative readings of the same sugar solution upon a polarimeter. The latter instrument does away with the errors of rotation dispersion and, aside from the objection of using monochromatic light, is always to be preferred in methods
same conditions
where the concentration or percentage of sugar is calculated from the If a quartz-wedge saccharimeter is the only instruangular rotation. ment available, the average factor 0.346 may be used for most purposes without serious error. Normal Weights of Sugars.
If a
sugar be taken for polarization, (i.e. dissolved to 100 c.c. will give a scale reading of 100), the percentage (uncorrected) of sugar may be read directly upon the saccharimeter.
normal weight of each particular the weight of pure sugar which
There are a number of methods of calculating the normal weight If we assume in case of the Ventzke scale that for different sugars.
the angular rotation of each division is 0.34657 circular degree for all sugars, then the normal weight (20 C., 100 true c.c.) of any sugar, for the 2-dm. observation tube, as compared with 26.00 gms., will be inversely proportional to the specific rotations of this sugar that is:
[a]g: 66.5
: :
and
of sucrose,
26 gms.
X, whence
(the
normal weight)
1729
f-W' a\D
\.
The normal weights of

in the following table:
several sugars calculated
by
this
method
are given
TABLE
XXXV
Giving Normal Weights of Different Sugars for Ventzke Scale

Sugar.
198
SUGAR ANALYSIS
While the normal weights calculated in
this
manner are
sufficiently
exact for most purposes of analysis they must not be regarded as absolute. Owing to the differences, previously mentioned, in rotation dispersion for the different sugars the angular rotation of each
Ventzke-scale division will vary slightly from 0.34657 circular degree with a corresponding change in the value of the normal weight. If the value of the 100-degree saccharimetric reading of each sugar
has been established in circular degrees, for the same conditions under which analyses are made, it is always better to base the calculation of
the normal weight upon this. The method of calculation for the Ventzke scale, using as illustrations four of the sugars previously taken, is as follows:
From
Glucose
(1
the general formula
IX
f-r
we obtain
=
l
for
[a\D
V.
0.3448 circular degree, Landolt),
X oo.4o
***' 4 * = 32.248 gms.

2 *J?* =
Lactose
(1
V.
0.3452 circular degree, Landolt)
32.857 gms.
Maltose
l- V.
. 0.3449
2
eircular degrees,
=
&
x\
,
'
12 , 74
gms
Raffinose + 5 H O
(1
V.
0.3450 circular degree, Landolt)
1 104.0
16.507 gms.
The conversion factors to be employed, and hence the values of the normal weights, will necessarily depend upon the quality of the light used for illuminating the saccharimeter. The value of a saccharimeter
division in circular degrees for a solution of the sugar of the
approximate
concentration, should, therefore, be established by the chemist himself
wherever possible.
Correction for Concentration and When normal Temperature. weights of the different sugars are used, the observed saccharimeter readings require correction for changes in concentration and temperature as described on page 195. Where much work is done with a single sugar a table of corrections should be prepared, giving the actual sugar value corresponding to each scale division of the saccharimeter. The correction table for sucrose (page 118) or the following results calcu-
by Browne* for glucose upon the basis of the normal weight of 32.25 gms. will illustrate the method.
lated
*
J.
Ind. Eng. Chem., 2, 526.

Scale division.
199
200
SUGAR ANALYSIS
1
on page 194 by the angular rotation of

scale (page 145), thus:
1
1
degree of the saccharimeter
angular rotation
D
= =
=
0.4785
0.4785 4785
. .
1 1
Ventzke sugar scale French sugar scale Wild sugar scale
X 0.34657 = 0.1658 gm. X 0.21666 = 0. 1037 gm. X 13284 = 0635 gm.

.
0.4785 gm. arabinose. arabinose. arabinose. arabinose.
Owing
to the lack of absolute agreement in the value of each sac-
charimeter scale in circular degrees, due to rotation dispersion, variation in quality of light, etc., the equivalent of 1 degree of a saccharimeter of the weight of sugar, which will give a scale is best expressed as T
reading of 100 degrees under the prescribed conditions of analysis (i.e. T The correction for concentration is afterwards of its normal weight).
applied as indicated above. The approximate value of 1
V. for the more
common
sugars
is
given below.
Weight of Sugar in 100 metric ex. V. 1 V. 1 V. 1 V. 1 V. 1 V. 1 V. 1 V. 1 V. 1 V.
1
at 20 at 20 at 20
at 20
C.
C. C.
C.
C. C. C. C. C.
at 20
at 20 at 20 at 20
at 20
at 20
C.
= 0.2600 gm. = 0.3225 gm. = 0.1859 gm. = 0.3286 gm. = 0.1247 gm. = 0.1655 gm. = 0.9100 gm. = 0.2135 gm. = 0.8645 gm. = 0.1651 gm.
sucrose.
glucose. fructose.
lactose hydrate. maltose.
arabinose.
xylose.
galactose. invert sugar.

raffinose hydrate.
For many laboratory Sugars. convenient to but fixed one normal purposes employ weight for all saccharimetric work. In such cases the normal weight of sucrose is usually taken, the percentage of each particular sugar being calculated from the scale reading by means of an appropriate factor.
of
it is
Use
One Normal Weight for All
polarizations in degrees Ventzke of a normal weight of different gms. sugars, when dissolved to 100 metric c.c. and in a 200-mm. The values polarized tube, are given in table
of 26.00
The constant
XXXVI.
are calculated only to the nearest 0.5 degree, which is sufficiently exact when the variations due to change in concentration are considered.
If no other optically active substances are present, the scale reading (V.) of 26.00 gms. of the sugar-containing substance multiplied by 100 and divided by the corresponding polarizing power of the pure sugar will give the percentage. of sugar present. Owing to the changes in specific rotation with varying concentration, the percentages thus calculated will not be absolutely exact.

TABLE
201
XXXVI
c.c.
Giving Ventzke Reading of 26.00 gms. of Different Sugars in 100
Sugar.
202
SUGAR ANALYSIS
perature, under the conditions specified above, provided that the sugar solution be made up to volume and polarized at this same temperature. "In effecting the polarization of substances containing sugar employ
only half-shade instruments. "During the observation keep the apparatus in a fixed position and so far removed from the source of light that the polarizing Nicol
is
not warmed.
"As sources of light employ lamps which give a strong illumination such as triple gas burner with metallic cylinder, lens and reflector; gas lamps with Auer (Welsbach) burner; electric lamp; petroleum duplex lamp; sodium light. "Before and after each set of observations the chemist must satisfy himself of the correct adjustment of his saccharimeter by means of
standardized quartz plates. He must also previously satisfy himself of the accuracy of his weights, polarization flasks, observation tubes and cover-glasses. (Scratched cover-glasses must not be used.) Make
several readings
and take the mean thereof but no one reading

,
may
be
neglected.
"In making a polarization use the whole normal weight for 100 or a multiple thereof, for any corresponding volume.
c.c.,
"As
clarifying
and decolorizing agents use
either subacetate of
lead, alumina cream, or concentrated solution of alum. Boneblack and decolorizing powders are to be excluded. "After bringing the solution exactly to the mark at the proper temperature, and after wiping out the neck of the flask with filter paper, pour all of the well-shaken clarified sugar solution on a rapidly
acting
filter.
Reject the
first
portions of the filtrate and use the rest,
which must be perfectly clear for polarization." Methods of the New York Sugar Trade Laboratory., Details of manipulation for the above rules are left largely to individual preference or requirement.
required,
is
The course of operations pursued by the New York Sugar Trade Laboratory, where rapidity as well as accuracy is
as follows:
Weighing. Twenty-six grams of sugar are weighed out in a nickel sugar dish provided with a counterpoise (Figs. 116 and 123). The
is stirred with a horn spoon and, approximately, the normal weight transferred to the dish. The final adjustment is then made with the dish upon the scale pan of the balance, a little sugar being added or removed until the exact weight is secured. The danger of spilling sugar upon the scale pan during the weighing is thus largely avoided. The weighing is performed as rapidly as possible to avoid loss from
sugar

minute of time.
203
evaporation of moisture and does not usually consume more than a
The 26 gms. of sugar in the nickel dish are poured Transferring. into a large funnel placed in a sugar flask; any sugar adhering to the dish and funnel is then washed into the flask with distilled water, the funnel being thoroughly rinsed inside and outside around the bottom
to insure the complete removal of all sugar to the flask. c.c. of water are sufficient to effect the transference.
From 50
to 60
funnels employed in transferring the sugar are of German and have a mouth 4 in. (ll cm.) in width and 3 in. (9 cm.) in silver, and a stem 3 in. (9 cm.) in length. The inner diameter of the depth,
The
(a)
(b)
(c)
Fig. 123.
(a) Nickel weighing dish and counterpoise. sugar, (c) Normal and half-normal metric
(6)
c.c.
Funnel for transferring

sugar weights.
stem (8J mm.)

into the flask
is
sufficiently large to allow a free passage of the sugar
and the outer diameter (10 mm.) sufficiently small allow the escape of air from the flask (see Fig. 123).
to
The solution of the sugar in the flasks is performed by Dissolving. means of a mechanical shaker. The machine employed in the New York Sugar Trade Laboratory is a modification by the author of the Camp shaker used in iron and steel laboratories. (Fig. -124.) The metal disk of this shaker is replaced by a circular piece of oak 1 in. thick, of the same diameter and of about the same weight, and containing 12 holes 2J in. in diameter, each large enough to accommodate the bottom of a sugar flask. Six extra gripping devices are inserted in
the collar of the shaker, thus giving 12 grips in all to hold the necks of the flasks. The collar is adjusted so as to bring the grips at the
right height and exactly over the centers of the circular holes in the wooden disk. The bottom of the flasks are inserted in the holes, and,
204
SUGAR ANALYSIS
the grips, the flasks are by pressing the necks against the springs of The shaker is connected snapped quickly and securely into position.
with a small J horse-power electric motor, provided with a rheostat, and the speed of its driving wheel gradually brought up to 120 to 130 revolutions per minute. At this speed, solution of sugar in the flasks, using 50 to 60 c.c. of water, is effected in from 5 to 10 minutes, according
to the size of grain, stickiness of sample, etc.
If
too
much water
body
is
used in transferring the liquid, and a longer time
sugar, less
is
motion
is
given to the
of the
required to effect solution.
Fig. 124.
Mechanical shaker for dissolving sugars.
The solution is then clarified with the requisite Clarifying. amount of lead subacetate solution (sp. gr. 1.25), but no more than the amount necessary to secure a clear polariscope reading is ever employed. As a rule not over 1 c.c. of the lead subacetate solution is used for Java, Peruvian, and high-grade centrifugal sugars, not over 1 to 2 c.c. for muscovado sugars, from 2 to 6 c.c. for molasses sugars, and 3, 4, and 5 c.c. for Philippine mat sugars according to grade. Excess of lead solution increases the polarization very markedly and
strict
observance
is
for clarification.
paid to the rule of minimum quantity necessary After the lead solution 2 c.c. of alumina cream are
added, the contents of the flask are well mixed and the volume of liquid made up to 100 c.c., after allowing sufficient time for any air bubbles
to arise which
may have
been occluded in the lead precipitate.
Foam
and
air bubbles,
the flask,
adhering to the surface of the liquid in the neck of are broken up with a fine spray of ether before adjusting the

volume to the graduation mark. is convenient for removing foam.
205
small bulb atomizer (Fig. 125)
The distilled water used in all the work is supplied through rubber tubing from a large bottle placed at an elevation above the laboratory table. The
outlets of the rubber tubes
are fitted with pinch cocks
and
glass tips of large
and
fine opening, the
former
being used for transferring the sugar and the latter for
The setting the meniscus. meniscus of the adjustment

to the graduation mark is the same as that used in
calibration (Fig. 119).

distilled
The
/\
I
water used for solu-
kept as nearly as o 2Q c> completion of the volume of sugar solution to 100 c.c. is always made with the contents of the flask at this temperature.
tion
Fig.
125.-Ether
atomizer.
^^ ^
is
^^
Filtering.
The contents of the flasks after thorough mixing
upon plaited filters in stemless funnels resting in All glassware is thoroughly or J-pint jars cylinders (Fig. 120). cleaned and dried before using. The plaited filters, which are large enough to hold the entire contents of the flask, are kept
are poured
a large desiccator until ready for use. The funnels are covered with watch glasses during the filtration to prevent concentration of liquid through evaporation. The first runnings (10 to 15 c.c.) of the filtrate are rejected and the rein
mainder used
for polarization.
Methods for Polarizing Juices, Sirups, Molasses,

Massecuites,
etc.
The method
of polarization just described for
raw sugars
juices,
may
be applied with minor modifications to the examination ^8- 126.

sugar-beet, sorghum,
of sugar-cane,
and other plant
sirups, molasses, massecuites,
and
all
other products which are
p i pe tte.
mostly soluble in water. Sucrose Pipette. In the analysis of sugar-containing juices the work of analysis may be lightened considerably by the use of Spencer's
206
SUGAR ANALYSIS
shown in Fig. 126. This pipette is gradwith stem the uated upon divisions, divided into tenths, reading from is so calibrated that the volume of juice deThe 5 to 25. pipette the division from livered upon the stem, which corresponds to its
or Crampton's sucrose pipette
degrees Brix, is exactly a double normal weight. The pipette is constructed either for Mohr cubic-centimeter or true cubic-centimeter
flasks,
The method
delivering 52.096 gms. and 52.000 gms. of juice respectively. of employing the pipette is thus described by Spencer.* " Determine the density of the juice with a Brix hydrometer,
noting the degree Brix without temperature correction. Fill the pipette with juice to the mark corresponding with its observed degree
Add 3 to 5 c.c. of diluted Brix, and discharge it into a 100-c.c. flask. lead-subacetate solution, complete the volume to 100 c.c. with water,
mix thoroughly and filter the contents of the flask. Polarize the filtrate, using a 200-mm. tube, and divide the polariscope reading by
2 to obtain the percentage of sucrose. The juice should not be .expelled from the pipette by blowing, and sufficient time should be allowed for
thorough drainage. Each pipette should be tested when received from the maker, and in regular work should be used under the conditions of the test. The pipette may be conveniently checked against a balance by delivering a measured quantity of juice into a tared capsule and
weighing it. The uncorrected degree Brix and juice of the temperature of the Brix observation must be used. If the hydrometer and pipette
are correct at the parts used, the juice delivered should weigh 52.096 gms. (or 52.00 gms. for true cubic centimeters). " It is not advisable to use these pipettes with liquids of a higher density than 25 degrees Brix or of greater viscosity than cane juice.
These pipettes are usually used in the analysis of miscellaneous samples
and in the rapid testing of diluted massecuites and molasses for guidance in the vacuum-pan work. They should be frequently cleaned with a strong solution of chromic acid in sulphuric acid."
of juice
For the analysis of highly concentrated sugar products, such as normal weight of substance is weighed out as with raw sugar. In case of very dark-colored molasses and massecuites, it is often necessary to make the normal weight of substance after clarification up to 200 c.c. instead of 100 Q.C. in order to reduce the depth of color sufficiently to polarize in a 200-mm. or, even at times, in a 100-mm. tube. The reading thus obtained is mulsirups, molasses, massecuites, etc., the
tiplied
by 2 (or if polarization is made in a 100-mm. tube by 4) to obtain the true direct polarization. * Spencer's "Handbook for Cane Sugar Manufacturers " (4th Ed.), p. 122.

CLARIFYING AGENTS AND ERRORS ATTENDING THEIR USE
207
In the clarification of dark-colored molasses and other sugar-house products a much larger amount of clarifying agent must be used than is necessary with raw sugars, juices, and other substances of high
of excessive quantities of clarifying agent serious errors in the work of polarization. These introduces, however, errors for convenience will be considered under the following heads:
purity.
I.
The employment
II.
Errors due to the volume of precipitated impurities. Errors due to precipitation of sugars from solution.
Errors due to change in specific rotation of sugars. influence of these errors will first be considered in connection
III.
The
with the different acetates of lead which are the salts most generally used for clarification.
Acetates of Lead. Three well characterized acetates of lead* have been isolated in the crystalline form. These are (1) the normal
,3 H 0; (2) the basic acetate Pb(C H H H the basic acetate Pb(C H O ,2 PbO, )2,PbO,3 Pb(C 0; (3) in general 4 H 0. The clarifying power of solutions of these acetates
or neutral acetate of lead

2 3 2 2
2) 2
2) 2
is
proportionate to the content of basic PbO. The normal acetate, although deficient in decolorizing power and unsuited for the clarification of darkthat
colored products for polariscopic readings, has certain advantages in it does not precipitate reducing sugars from solution and does not
form soluble lead-sugar compounds of different specific rotation. For these reasons the neutral acetate of lead should be employed for clarifying wherever possible in preference to the basic salt. Neutral Lead-acetate Solution. In preparing the neutral acetate of lead reagent, a concentrated solution of commercial lead acetate
(sugar of lead) is made, any free alkali or acid neutralized with acetic acid or sodium hydroxide, and the liquid diluted to a density of 30 The solution is degrees Be\ (54.3 degrees Brix or 1.2536 sp. gr. ^).
filtered
and kept in a stock bottle ready for use. Lead-subacetate Solution. Upon digesting litharge with normal
acetate of lead solution varying amounts of lead oxide are dissolved acNumerous methods cording to the time and temperature of digestion.
are
employed
for preparing lead-subacetate reagent.
The
following
examples are given: I. Concentrated Solution.]

*
Heat, nearly to boiling, for about half

litharge,
an hour, 860 gms. of neutral lead acetate, 260 gms. of

R. F. Jackson:
"
t Spencer's
and
Handbook
Scientific Paper, for Cane
U. S. Bureau of Standards, No. 232 (1914). Sugar Manufacturers," p. 229.
208
500
c.c.
SUGAR ANALYSIS
of water.
Add water to compensate for the

clear solution.
loss
by evaporation.
may be prethe mixture is set without aside several hours heat, provided pared with frequent shaking. Proceed as described above, using, however, Dilute Solution. 1000 c.c. of water. The solution should be diluted with cold, reCool, settle,
solution
and decant the
The
degrees
cently boiled distilled water to 54.3 Brix (30 degrees Be., or
1.2536 sp.gr. ^). II.* Boil 430 gms. of normal lead

acetate, 130 gms. of litharge, and 1000 c.c. of water for half an hour.
Allow the mixture to cool and
settle
and
dilute the
to 1.25 sp. gr. distilled water.

Ill.t
supernatant liquid with recently boiled
lead acetate
Treat 600 gms. of neutral and 200 gms. of litharge

After stand-
with 2000 c.c. of water.
ing 12 hours in a warm place with occasional shaking, the solution is

filtered
and the
nitrate
stored
in
tightly stoppered bottles.
tion
The soluthus prepared must show a
strongly alkaline reaction and have specific gravity of 1.20 to 1.25 (at
17.5
C.) with a content of about 20
per cent PbO. IV. Lead - subacetate

also
solution
be prepared by dissolving may the solid basic salt (see page 214). The concentrated solution is diluted with distilled water to a specific
Fig. 127.
Stock bottle and burette for
gravity of 1.25.
lead subacetate solution.
Stock solutions of lead subacetate,
both in bottle and burette,
should be protected by a soda-lime tube from the carbon dioxide of the air to prevent deposition of lead carbonate (see Fig. 127).
* "
Methods
of Analysis A. O. A. C.," Bull. 107 (revised), U. S. Bur. of Chem.,
p. 40.
t Fruhling's
"Anleitung," p. 457.

I.
209
Errors of Clarification
Due
to
Volume
of Precipitated Impurities
Since all sugar solutions after clarification with lead subacetate, or other means, are made up to a definite volume, the space occupied by the precipitated impurities will cause the sugar solution to occupy a somewhat smaller volume than that of the flask in which the solution
was made up.

is
An
increase in concentration
and
also in polarization
the result.
Scheibler's Method of Double Dilution. Several methods have been devised for estimating the extent of this error. The first to be described is Scheibler's* method of double dilution. In this method a normal weight of product is dissolved in water, clarified with a measured volume of lead subacetate, the volume completed, and solution A second normal weight of product filtered and read in the usual way. is then weighed out, clarified with the same volume of reagent as before
and the solution made up to twice the volume of the previous experiment. The second solution is filtered and polarized as before. The true polarization (P) is then calculated as follows: Let PI be the polarization of the first solution made up to volume
V, and P% the polarization of the second solution made up to volume 2 V. Let v be the volume of the precipitated impurities which is assumed to be the same in both experiments. The normal weight in
the second solution

half dissolved in
may
volume
be considered to be divided as follows: one free from precipitate, the reading of which
would be
p
j
and one
half dissolved in
volume
containing precipitate,
the reading of which would be -^ 2
The sum
of these quantities divided
by 2
is
the value of
2,
or
= 4 2 PI. In other words the true polarization is equal whence to four times the polarization of the diluted solution less the polarization of the undiluted solution, f
Deut. Zuckerind., 25, 1054. true polarization is also expressed in other ways as: multiply reading of dilute solution by 2, subtract the product from reading of undiluted solution; twice the remainder subtracted from reading of undiluted solution will give the true
Z. Ver.
f
The
polarization: or the difference between the reading of the undiluted solution, and twice the reading of diluted solution subtracted from twice the reading of the diluted solution will give the true polarization.
210
SUGAR ANALYSIS
Polarization of 26 gms. raw sugar, dissolved in water, clarified Example. with 2 c.c. lead subacetate and made to 100 c.c. = 94.2 (Pi). Polarization of 26 gms. same sugar, dissolved in water, clarified with 2 c.c.
lead subacetate
and made to 200
c.c.
True polarization (P)
(47.0
4)
47.0 (P 2 ). 94.2 = 93.8.
The volume
as follows.
occupied by the precipitated impurities

of the undiluted solution
is
is
calculated
The reading PI
(Pi
equal to
whence
= V
VXP y_
>
P)
r\
Example.
example.
Substituting the values for V,
9
Required the volume of the lead precipitate hi the previous
P and Pi,- we obtain

93 8)
'
10
(94
'V2
its
42
c c-
The method
of Scheibler
owing to
rapidity
and ease
of execution
has been very widely used for correcting polarizations for the error due to volume of the lead precipitate. The method is open to several
objections.
It is not probable that the volume of the precipitate is the in the dilute as in the undiluted solution, but the prinsame exactly cipal objection against the method is the very large multiplication of
any error made

Sachs's
devised
in reading the diluted solution.
The method of Correcting Precipitate Error. in Sachs* due to volume of 1880 for the error by determining method. was intended to Scheibler's obviate the errors of precipitate In the Sachs method the precipitate of impurities obtained in the
clarification of the sugar solution is
Method
until all
washed with cold and hot water removed. The sugar precipitate is then transferred to a 100-c.c. flas'k, a one-half normal weight of sucrose added, the latter dissolved and the volume completed to 100 c.c. The solution is mixed,
is
filtered,
is
and polarized in a 400-mm. tube. The volume of precipitate then calculated as follows: Let P = the true polarization of the sucrose used and PI = the polarization of the sucrose with precipitate.
The volume
(v)
of precipitate
is
then found by the equation
100 (Pi
P)
Pi
Example.
normal weight
of granulated sugar dissolved to 100 c.c.
polarized 99.8 in a 200-mm. tube. A one-half normal weight of the same sugar to 100 c.c. polarized 100.25 in a 400-mm. tube.
-+-
lead precipitate dissolved

of precipitate (v)
Volume

Knowing the volume
of a product
(v)
211
of lead precipitate, the true polarization (P)
may
be determined by the equation
P=
or
when
V=
100,
P=
~*;Pl
.
^Q
Example.
96.20 (Pi).
The polarization of a raw sugar (26 gms. to 100 c.c.) was The volume of the lead precipitate bySachs's method was 0.22 c.c. (v).
The
true polarization (P) of the sugar
100
96.2
0.22
96.2
95.99.
100
The method of Sachs has been modified as follows. Instead of making a polarization with the washed precipitate the latter is first dried. From the weight and specific gravity of the dried lead precipitate the
volume
is
calculated [v
=
sp. gr.
and from the volume the
determined by means of the preceding formula. specific gravity of the dried lead precipitates of raw cane sugars was determined by Wiechmann* by weighing in a pycnometer with benzine. The results of Wiechmann are given in Table XXXVII.
true polarization
is
The
TABLE XXXVII
Giving Specific Gravity and Volume of Lead Precipitates from 26 gms. of Different
Raw Cane
Sugar.
Sugars
212
SUGAR ANALYSIS
work
of clarification; they are not adapted, however, to practical owing to the large amount of time and labor involved.
Home's Method
ing the
of
Dry Defecation.
A third method of eliminat-
volume of precipitate error is Home's* process of dry defecation. The method is thus described by its author: " The normal weight of sugar is dissolved in water in a 100-c.c. The concentraflask and made up to the mark without defecation.
It now remains to defecate tion is thus at exactly the proper degree. the solution properly by precipitating the impurities in such a way as to produce the minimum change in the concentration of the solution of
sucrose.
This
is
accomplished by adding to the 100
c.c.
of liquid
small quantities of powdered anhydrous lead subacetate until the imThis point is as easily determined purities are nearly all precipitated.
as in the defecation by a solution of the same salt. The organic and mineral-acid radicals in the solution combine with and precipitate the lead and lead oxide of the dry salt, while the acetic-acid radical of the lead subacetate passes into solution to combine with the bases originally united to the other acid radicals."
12 raw cane sugars are given in a very close agreement between the corrected polarization by Sachs's method and the polarization by dry
Results obtained by
Home upon
Table
XXXVIII, and show
defecation.
TABLE XXXVIII

ever, has criticized the increase in polarization
213
principally upon the ground that the due to the volume of precipitate is not as great as calculated, owing to the decrease in polarization caused by the
method
retention of sucrose in the precipitate, this retention error frequently more than counterbalancing the error due to volume of precipitate.
Subsequent results by Home* and other chemists show, however, that is no appreciable retention of sucrose when the dry lead reagent is used in minimum amounts. Another objection by Pellet, that only
there
part of the lead salt acts and that the rest passes into solution, thus increasing the volume and diminishing the polarization, deserves consideration.
of sugar-house products there is no difficulty a satisfactory clarification with a minimum amount of the dry lead salt, the lead dissolved being immediately precipitated and but very little remaining in solution. With low-grade sugars, molasses, If dry lead subacetate, or subacetate soluetc., the case is otherwise. tion, be added to a solution of such products to the point of satisfactory clarification a considerable amount of lead salt will usually remain dissolved. The rule of adding the powdered salt until no more
in securing
With the higher grade
not always a criterion of the absence of lead in the is added to solutions of low purity the first portions of lead are completely precipitated; then comes a point where with the formation of additional precipitate a small amount of lead
is
precipitate forms
filtrate.
When
subacetate
remains in solution; the amount of the latter continues to increase until at the point where no more precipitate is formed nearly all of the lead added remains dissolved. With very low (See Table XXXIX.)
grade products there is therefore a danger of the dry lead salt increasing the volume of solution; whether this increase will cause a lowering of
the polarization or not will depend upon the character of the product. With low-grade sugar-cane products the error due to increase in volume
of
solution
may
be more than counterbalanced by the precipitation
of levorotatory fructose.
In the following experiments by Hall f in the New York Sugar Trade effect of increasing amounts of dry lead subacetate upon the polarization of a Philippine mat sugar was studied. The quantity
Laboratory the
of lead in the clarified filtrates
lated
was determined and the dilution calcuby allowing an increase of 0.22 c.c. in volume for 1 gm. of dry
c.c.
subacetate dissolved in 100

*
J.
of solution.
Soc., 29, 926. Bur. of Chem., p. 225.
Am. Chem.
U.
t Bull. 122,
S.
214
SUGAR ANALYSIS
TABLE
XXXIX
Showing Estimated Dilution of a Sugar Solution by Dry Lead Subacetate

of the basic acetate 3
215
Pb(C 2 H 3
PbO.*
)2,PbO and three parts of the basic
acetate
Pb(C H
2
2 ) 2 ,2
solution of lead subacetate of 1.259 sp. gr., as employed for clarification in the wet way, was found to contain 0.2426 gm. total Pb per
Ic.c.
One-third gram dry salt
is
therefore equivalent to
1 c.c.
subace-
tate solution in clarifying power. low-grade sugar requiring 6 c.c. of subacetate solution of the above strength for clarification would
accordingly need 2 gms. of salt for dry defecation. The dry subacetate of lead employed in sugar analysis should be finely ground in order that it may be acted upon quickly and comThe tendency to form insoluble pletely by the dissolved impurities.
crusts
upon the powdered grains
of dry salt has been noted
by Home,
especially in refinery products subjected to the influence of
In such cases
Home recommends
the addition of a
little
bone black. dry sand with

off
the powdered lead salt; the particles of sand in shaking will grind the crusts of insoluble matter and allow the lead to be acted upon.
II.
Errors of Clarification due
to
Precipitation of Sugars
from Solution
In the absence of free alkalies sucrose

tion
by lead subacetate.
is not precipitated from soluReducing sugars, however, are precipitated
by
solutions of basic lead salts.
This precipitation does not occur
with the amounts of lead used in ordinary clarification except in presence of those salts or acids which form insoluble lead compounds! (as
sulphates, phosphates, carbonates, oxalates, tartrates, Whether this precipitation of reducing sugars is due malates, etc.). to simple occlusion or to the formation of insoluble sugar-lead comchlorides,
plexes
not definitely known. to which the common reducing sugars, glucose and fructose, are precipitated by different lead clarifying agents, has been
is
The extent
investigated by Bryan. J Separate solutions of glucose and fructose were prepared, using 5 gms. of sugar with 1 gm. each of magnesium sulphate and ammonium tartrate. To 50 c.c. of this solution the After clarifying agent was added and the volume made up to 100 c.c. filtering, the excess of lead was removed with potassium oxalate, and the sugar in solution determined by Allihn's method. The results of
Bryan's experiments are given in the following table.

*
Home's dry subacetate

t
t
Jackson in an unpublished experiment communicated to the author shows that is in fact a mixture of these two basic acetates. Prinsen Geerligs, Deut. Zuckerind., 23, 1753. Bull. 116, U. S. Bur. of Chem., p. 73.
216
SUGAR ANALYSIS
TABLE
XL
Showing Precipitation of Glucose and Fructose by Basic Lead Salts
Clarifying agent.

TABLE XLI
Number of cubic centimeters of basic lead solution
(1.25 sp. gr.)
217
added.
218
should be added to
clarified solution to
SUGAR ANALYSIS
weak
100
acidity before making up the volume of the for the direct polarization of low-grade
c.c.
fructose containing products.
Miscellaneous Methods of Clarification
Numerous
modifications of the lead process of clarification have
been proposed as a means of reducing or eliminating the several sources of error just mentioned. Freshly precipitated lead carbonate, lead chloride, and lead nitrate have been employed as clarifying agents, but
Two methods of lead clarification, with only indifferent success. which have found considerable favor in France and Austria, should, however, be mentioned in addition to the processes previously described.
These are Zamaron's method by means
of hypochlorite of of basic
lime and neutral lead acetate, lead nitrate.
and Herles's method by means

Clarification with
Zamaron's
grams
of
Method
of
Hypochlorite.
625
dry commercial bleaching powder are thoroughly ground up The mass is squeezed out in a large mortar with 1000 c.c. of water. The solution thus in a sack and the extract filtered through paper. obtained (700 c.c. to 800 c.c. of about 18 Be.), is preserved in a stoppered bottle of dark glass away from the light. The solution to be clarified is treated with a few cubic centimeters
of the hypochlorite solution,
sufficient
to effect decolorization, and
then a few cubic centimeters of neutral lead acetate solution are added.
There
usually a slight rise in temperature after addition of the clarifying agents so that the solution must be recooled before making to
is
volume.
process secures usually a good clarification, does not precipitate reducing sugars, and forms no objectionable lead sugar compounds. The chief fault of the method is the volume of precipitate
error,
The Zamaron
which in this case
is
augmented by the formation
of considerable
lead chloride.
Herles's f
solve 100
Method
of solid
of Clarification with Basic
Lead
Nitrate.
Dis-
sodium hydroxide in 2000 c.c. of water; a second grams solution is prepared by dissolving 1000 gms. of neutral lead nitrate in 2000 c.c. of water. Upon mixing equal volumes of the two solutions
basic lead nitrate
is
precipitated according to the equation
Pb(N0 3 ) 2
nitrate
+ 2 NaOH
= Pb(NO
3)2
.Pb(OH) 2
+ 2 NaN0
Lead
Basic lead nitrate
Fribourg's "Analyse chimique," p. 129.
t Z. Zuckerind.,
Bohmen,
13, 559; 14, 343; 21, 189.

The precipitated basic lead nitrate is washed pounds and then mixed with water to a cream,
used for clarification.
free
219
in
from sodium comwhich form it may be
The
clarification
is
nitrate within the solution to be clarified.
performed more commonly by forming the basic This is done by first adding
a measured quantity of the lead-nitrate solution (1 c.c. to 15 c.c. according to depth of color) and then, after mixing, an equal volume of the so-
dium hydroxide solution. well mixed, and filtered.

solution
After shaking, the solution is made to volume, Care must be taken that the reaction of the
is not alkaline after mixing; this is best provided for by testing the two solutions against one another before using. Formation of the basic lead nitrate within the solution gives usually
a much better clarification than addition of the washed cream, but has the disadvantage of introducing considerable sodium nitrate, which, if present in large quantity, will affect the rotation of the sugars.
method gives an exceedingly brilliant clariopen, however, to the same errors as basic There is first the volume of precipitate error, which is lead acetate. further augmented by the copious bulk of the basic lead nitrate itself;
basic lead nitrate
fication.
The
The
process
is
and secondly there

the results of
is
a precipitation of reducing sugars as shown by

of basic lead
Bryan in Table XL. The numerous errors incident to the use
compounds
in clarification
have led chemists to seek other means
of decolorizing
solutions for polarization. It is impossible, as well as unnecessary, to take up all the processes which have been devised to accomplish this end. Two of these methods, however, should be described: (1) Decolorization
tion
by means of bone black or blood charcoal; (2) Decolorizaby means of hydrosulphites, sulphoxylates, etc. Decolorization of Sugar Solutions by means of Bone Black. The
use of bone black as a decolorizing agent in sugar refineries is well known. The same substance in a more finely divided specially prepared form is employed at times as a decolorizer in sugar analysis.
Purification of
Bone Black.
If purified
animal charcoal (preferably
blood charcoal) has not been obtained from the dealer the chemist may purify the commercial product as follows: The char is finely ground in a mortar and then digested several hours in the cold with dilute hydro-
then decanted, the char brought upon a water until all traces of hydrochloric acid are removed. After drying in a hot-air oven, the char is heated to dull redness in a covered porcelain crucible, and then, after cooling suffichloric acid.
filter
The
acid
is
and washed with
distilled
ciently, placed while still
warm
in
a dry stoppered bottle.
220
Several
SUGAR ANALYSIS
methods are followed
in the employment of animal charcoal common One for decolorizing. practice is to make up the soluvery tion to volume and shake thoroughly with a small quantity of charcoal, using from 0.5 to 3 gms. according to depth of color. The contents of the flask are then poured upon a dry filter and the filtrate taken for
polarization.
In the above method of decolorAbsorption Error of Bone Black. is introduced owing to the absorption and retenerror certain a izing, tion of sugar by the char. Sugars differ markedly in the extent to In the case of the simple the error through absorption is reducing sugars, glucose, fructose, etc., so small as to be almost negligible, but in the case of sucrose and other higher saccharides the absorption is so great that an error of several
which they are absorbed by animal charcoal.
degrees Ventzke
may
be occasioned in the polarization.
One method
of eliminating the error
through absorption of sucrose
consists in adding a correction previously established by experiment upon pure sugar solutions. If, for example, a sucrose solution polariz-
ing 95.0 V. gives, after shaking 50
c.c.
with 2 gms. of charcoal for 5
minutes, a polarization of only 94.7 V., then a correction of 0.3 V. must be added to all polarizations of about 95 V. for sugars decolorized in this same way. A correction table is thus made for sugar solutions of
but in applying these corrections care must be taken that the quality and quantity of the char are alike in both instances and that the time of shaking is always the same. With impure products of variable composition the employment of absorption factors is attended with considerable uncertainty.
different concentrations,
Spencer* has recommended a different method of employing animal

charcoal for the purpose of reducing the absorption error to a minimum. The process is thus described: " Place a small quantity of bone black, about 3 gms., in a small
Add a volume of plain filter, selecting a rather slow filtering paper. the solution equal to that of the char, or just completely moisten the
latter,
and
let this liquid filter off.
After four or five similar nitrations,
the filtrates from which are rejected, test the filtrates by a polariscopic observation and note whether the reading varies. Solutions must be protected from evaporation during the filtration. As soon as the reading is constant, showing no further absorption, record it as the required
number."
The method just described, while largely eliminating, does not completely remove, the errors of absorption, for while the retention of
*
Spencer's
"
Handbook
for
Cane Sugar Manufacturers"
(4th Ed.), p. 89.

sucrose
221
by the char rapidly diminishes with each

it
successive portion of
soon becomes only a gradually receding quantity. This is shown by the following experiments upon a sucrose solution polarizing
solution,
49.9
V.
Fraction of
filtrate.
222
SUGAR ANALYSIS
for bleaching dark-colored massecuites
where they have been used

also, in solution, as a
and
wash
for whitening sugars in the centrifugal.
They have
also
been employed by unscrupulous manufacturers for

is
bleaching low-grade molasses in the preparation of table sirups. For their use in sugar analysis the solution to be decolorized
treated with a lew cubic centimeters of alumina
cream and a few
crystals of sodium hydrosulphite (0.1 gm. to 1.0 gm., according to the depth of color) ; after mixing and dissolving, the volume is made up to
the mark, and the solution filtered.
The
filtrate
should be polarized
tjiere is a rapid redarkening of solutions decolorized with hydrosulphites. Weisberg,* from his study of the action of hydrosulphites, concludes that the bleaching action is a double one, first, by means of the free sulphurous acid when decolorization is per-
immediately. In many cases
manent, and secondly by means of the nascent hydrogen which is evolved, when there is a redarkening of the solution through oxidationAfterdarkening may be prevented by the use of another hydrosulphite derivative, sodium sulphoxylate-formaldehyde, sold commercially as
Rongalite." The latter, however, is much slower in its bleaching action than hydrosulphite and is not always an effective decolorizing
agent.
serious objection against hydrosulphite is its action upon the polarizing power of certain reducing sugars. Bryan f has found that the polarizing power of glucose was decidedly lowered after the addition of hydrosulphite, owing to the formation of a levorotatory oxy-
"
sulphonate. Rongalite did not produce this effect. Neither rongalite nor hydrosulphite caused any immediate change in the polarization of fructose or sucrose. Numerous cases of inversion of sucrose by the
prolonged action of hydrosulphites have been reported, however, in the

literature.
The experience
of chemists, in the use of hydrosulphites as a de-
colorizing agent for sugar analysis, has been upon the whole unfavorable. In many cases the decolorized solution becomes turbid through
separation of sulphur, thus rendering polarization impossible. bleaching action of hydrosulphite is also limited, and has but
decolorizing effect upon caramel substances, which are chief causes of discoloration in sugar-house products.
The
little
among
the
Aluminum Hydroxide as a
*
Clarifying Agent.
A common prepahaving but
ration, used in connection with other clarifying agents, yet

Centrbl. Zuckerind, 15, 975.
t Bull. 116,
U.
S.
Bur. of Chem., p. 76.
223
little decolorizing power in itself, is aluminum hydroxide, or, as it is more generally termed, "alumina cream." The method of preparing alumina cream, as prescribed by the Association of Official Agricultural
Chemists,
is
as follows:*
"Prepare a cold saturated solution of alum in water and divide Add a slight excess of ammonium hydroxinto two unequal portions. ide to the larger portion and then add by degrees the remaining alum solution until a faintly acid reaction is secured." The reagent as above prepared consists of aluminum hydroxide
suspended in a solution of ammonium and potassium sulphates. The have a certain advantage, when alumina cream is used as an adjunct with lead salts, in helping to precipitate any excess of lead from solution. In certain cases, however, the presence of ammonium
salts
and potassium sulphates is detrimental, so that for many purposes it is For the preparation of the latter, better to employ a salt-free cream. concentrated alum solution is precipitated with a slight excess of ammonia and then washed by decantation with water until the solution The excess of water is then poured off and is free from sulphates. the residual cream stored in a stoppered bottle. The clarifying effect of alumina cream is chiefly mechanical; its
action consists largely in carrying
down
finely
suspended or colloidal
When used in impurities which would otherwise escape filtration. connection with lead subacetate it promotes the coagulation of the
and renders filtration more perfect and rapid. For the polarization of very high grade sugars, sirups, honeys, etc., In all such cases alumina cream is the only clarifying agent required. 2 c.c. of the cream should be used. About only the salt-free reagent are sufficient for clarification and the volume of aluminum hydroxide
precipitated impurities
in this
amount
is
Concentrated alum solution

for clarifying.
too insignificant to affect the polarization. is sometimes used with lead subacetate
The precipitate, formed between the lead salt and alum, remove to helps coloring matter, but the increase in precipitate and other errors tend to nullify any advantages of the method.
Comparisons of Different Clarifying Agents.
A few examples, taken from the reports of Referees upon Sugar for the Association of Official Agricultural Chemists, are given in order to show the probable error of different clarifying agents in polarization.
*
Methods
of Analysis A. O. A. C. Bull. 107 (revised),
U.
S.
Bur. of Chem.,
p. 40.
224
SUGAR ANALYSIS
TABLE XLII
Polarization of Mixtures of Sucrose, Glucose, 0.5 gm. Ammonium Oxalate and 0.5 gm.
and Fructose with Sodium Sulphate,

*
using Different Clarifying Agents (Bryan)

Clarifying agent.
225
acetate and nitrate solutions give much higher polarizations owing to both the volume of precipitate error and the precipitation of fructose. Neutral
lead acetate solution and dry lead subacetate give polarizations between these two extremes, there being, however, in case of the former, a volume of precipitate error and in case of the dry lead an error due to precipitation of reducing sugars. The true polarization would be somewhere between the results obtained with hydrosulphite and neutral lead acetate. The selection of an appropriate clarifying agent is one of the most important operations of saccharimetry, and in making his selection the chemist must be governed by the requirements of each particular case. Rapid nitration and brightness of clarification are factors which must be considered as well as minimum degree of error. Beginning with products of highest purity alumina cream alone should be used wherever
possible.
is
With products
of slight discoloration,
when alumina cream
When insufficient, neutral lead acetate solution should be tried. alumina cream and neutral lead solution fail, lead subacetate, or basic lead nitrate, or neutral lead acetate with hypochlorite may be employed;
dry lead subacetate will usually give more accurate results with sugarcane and other products containing fructose. Animal charcoal or hydrosulphites should be used only as a last resort, when other means of clarification have failed. The smallest possible quantity of clarifying in should be used all cases. agent
POLARIZATION OF SUGAR PRODUCTS CONTAINING INSOLUBLE
MATTER
In the analysis of juices, sirups, molasses, massecuites, and sugars, the chemist has to deal with substances which are entirely soluble in
water.
of polarization becomes more complicated when considerable insoluble matter is present, as happens in the analysis of
fruits, tubers, stalks, and other vegetable substances or in the examination of filter-press cake, scums, and other sugar-house residues.
The work
The methods for polarization of succulent plant materials may be divided into three general classes: (1) Methods of Expression; (2)
Methods
of Extraction,
tration of these several
and (3) Methods of Digestion. As an methods the polarization of sugar beets
illus-
offers
a good and
classic
example.
In preparing sugar-beets, sugar cane, fruits, etc., for analysis the material must first be reduced to a For this purpose any of the numerous finely divided condition.
Sampling Sugar Beets, Etc.
mechanical rasps, shredders, graters, etc., may be employed, provided that the cellular tissue be thoroughly disintegrated and that no losses
occur through leakage of juice or evaporation.
226
Keil's Beet Sampler.
SUGAR ANALYSIS
The Keil boring machine (Fig. 128) is very for used taking samples of individual sugar beets. The frequently essential feature of the apparatus consists of a hollow detachable bit,
the construction of which
is
shown
in Fig. 129.
The
conical rasp at
Fig. 128.
Keil's boring rasp for
sampling sugar beets.
the end, revolving at a speed of about 3000 revolutions per minute, reduces the substance of the beet to an extreme degree of fineness and at
the same time forces the pulp through a small opening into the cavity
Fig. 129.
Detachable bit of Keil's boring rasp.
within.
Each beet
is
bored in an inclined direction, as shown in Fig.
When only 130, in order to secure the best representative sample. " " for mother beets single beets are examined (as in the selection of
seed production) the bit is detached after each boring and a new one screwed on. The bits are numbered, and to obtain the sample the conical rasp is removed and the pulp (from 8 to 14 gms., according to
the size of beet and length of boring) forced out with a rod. In sampling large numbers of beets the bit is kept in constant use, the pulp
227
being discharged in a continuous stream into a covered container at the end of the apparatus.
/.
Determination of Sugar in Sugar Beets by Expression of Juice

of the
of the sugar in sugar beets by polarization was expressed juice formerly quite common, but has now more to accurate methods of analysis. given place
The determination
Assuming
(as
is
incorrect) that
the sugar, amides,
albuminoids, salts, gums, and other water-soluble solids of the beet are in the same condition of solution within = the the beet as in the expressed juice, and letting " " cent of water-insoluble matter or marc and 100 per
the per cent of juice, then the sugar content (S) can be calculated from the polarization (P) of the expressed juice by the formula
of the beet
P(100
- M)
100
The expressed juice of a sugar beet gave a Example. polarization of 16.2 V. for the normal weight: the beet contained 4.6 per cent of marc. Required the per cent of sugar in
the beet.
16.2 (100
- 4.6)
100
15.45 per cent.
The above method is, of course, equally applicable to the analysis of sugar cane, fruits, and other succulent plant substances.
?'
rection
of in
boring
juice
of Expressing Juice. For expressing the sampling sugar beets. from the pulp of sugar beets, sugar cane, etc., any suitable form of hand press may be used. The small hydraulic press shown in Fig. 131 is one of great efficiency and is a piece of apparatus
Method
almost indispensable in a sugar laboratory. The pulp to be pressed is placed in a strong sack inside the perforated container C, and covered evenly with a heavy metal disk. By
the screw A is driven downward as far as possible turning the wheel upon the disk, thus squeezing out through the openings of C a considerable part of the juice, which escapes by the spout into a can or
other receptacle. The horizontal hydraulic screw B is then turned inwards. This screw, operating by means of glycerol which fills the
hollow base H, forces the piston upwards and removes by vertical The final pressure, indicated by pressure a second fraction of juice.
228
the manometer
SUGAR ANALYSIS
M, can be raised
to 300 atmospheres.
The
juice, as
the
important pressure increases, is of gradually diminishing purity; therefore that all the runnings should be well mixed before taking the
sample for polarization.
it is
Fig. 131.
Laboratory hydraulic press
for expressing juices.
determination of the insoluble cellular matter, or marc, is necessary before the per cent of sugar in plant substances can be calculated from the polarization of the expressed juice.
Determination of Marc.
For rough purposes of estimation a constant percentage of 5 per cent or 4.75 per cent marc is sometimes assumed for the sugar beet and 10
Such figures, however, per cent or 12 per cent for the sugar cane. matter varies conof cellular the have no exact value, as percentage of the the to plant, dryness of the season, and age siderably according other conditions. many
For the determination of marc 20 to 50 gms. of the finely divided pulp are digested with 200 to 500 c.c. of cold water for 30 minutes, and then filtered as dry as possible upon a piece of finely woven linen, using suction. The washing is repeated with successive portions of cold water until the filtrate, from color and taste, is judged to be free of extractive matter. The residue is then washed several times with hot distilled water, then, after pressing together, with 2 to 3 portions of

90 per cent alcohol, and
volatilized the
229
After the ether has finally with a little ether. dried in an oven, gradually raising the temperature after a few hours to between 100 and 110 C. After cooling in a desiccator the residue, which is very hygroscopic, is rapidly
marc
is
weighed
(preferably in a stoppered weighing bottle) and the weight taken as the amount of cellular matter or marc. For a determination of the
organic cellular matter, the marc
is
incinerated
and the percentage
of
ash deducted.
The percentage
of juice.
of
marc subtracted from 100 gives the percentage
Where many determinations of marc have to be performed, a battery of small continuously operating percolators will effect a conMethod. Several sources of error are involved in the determination of sugar in plant substances by analysis of the expressed juice. In the first place a considerable amount of
juice,
siderable saving of time. Errors of Expression
ciency of the press,
varying from 10 per cent to 30 per cent, according to the effiis not eliminated and this residual juice, containing
a larger amount of albuminoids, pectin, etc., is of much lower purity than the part first expressed. This excess of impurities in the unexis washed out, however, in the marc determination. polarization of the expressed juice is thus higher than that of the composite juice of the entire plant. (See under Distribution of Water,
pressed juice
The
page 230.)
is the extraction during the marc determiamounts of cold water, but more especially of variable amounts of hemiby the hot water, alcohol, and ether celluloses, wax, oil, and other substances which are, strictly speaking, not juice constituents and should therefore be included in the marc. The percentage of juice is thus estimated too high, and a plus error
second source of error
nation
by the
excessive
introduced in the calculation. Except for the disadvantage of loss of time in drying, the use of alcohol and ether as dehydrating agents should be omitted in the marc determination, and cold water alone be used for extracting.
"
Colloidal
"
or
"
Imbibition
"
Water.
be mentioned is the much-debated tion" water, by which is meant
question of water, in a
"
third source of error to " " imbibior colloidal
more or
less
hydrated
form, in combination with hemicelluloses and other plant constituents. This imbibed water contains no sugar in solution, and, being expelled from the pulp upon drying, the percentage of sugar-containing juice is
overestimated.
230
SUGAR ANALYSIS
Heintz* showed, in 1874, when the air-dried and sugar-free marc of beets was placed in sugar solutions, that water was imbibed, thus leavIn ing the sugar more concentrated and increasing the polarization.
air-dried beet marc, which had from been washed sucrose, was treated 16 hours in a solutions with cool place containing a normal and half-normal weight in the of sucrose, proportion of 1 gm. marc to 20 c.c. of solution.
the following experiments
by Heintz
completely free

Fig. 132
stalk.
231
shows a magnified cross section of a part of a sugar-cane
juice proper, represented by S (the vacuoles), constitutes the principal part of the cell contents in the thin-walled parenchyma or fundamental tissue, and includes the greatest part of the
The sugar-containing
water in the cane.
Lining the walls and permeating
Fig. 132.
Magnified cross-section of sugar-cane (protoplasmic

lining
P much
intensified)
through these cells are thin layers and threads of protoplasmic matter which contains a considerable amount of water, but is deficient in
sugar.
fibro vascular
Running longitudinally through the stalk are large numbers of bundles whose ducts, D, are filled with water taken up
from the soil. The water of these ducts may often be seen spurting from the end of a cane stalk as it passes between the rollers of a mill, and is found upon analysis to be almost free of sugar. Running parallel
T which carry in solution the products of assimilation from the leaf to the stalk. The water of these
with the ducts are the sieve tubes
tubes contains reducing sugars but is deficient in sucrose. The cellular walls of the parenchyma and fibrovascular tissues contain about 50
per cent cellulose, 20 per cent xylan, 5 per cent araban and a remainder of lignin substances, all of which may hold a certain amount of water in the imbibed or colloidal form.
232
SUGAR ANALYSIS
The pressVariation in Composition of Juice from Different Mills. mill consist mostly of cane of a or crusher rollers first the from ings the sugar-containing juice S (Fig. 132). The pressings from succeeding rollers, where the pressure is greater, contain more and more of the protoplasmic juice P and the juice from the ducts and tubes. The colloidal water of the cellular substance is of course not affected by
the milling.
The composition of the pressings from the different cane mill is given in Table XLIV.
TABLE XLIV
rollers of a

juices,
233
faulty
marc estimation, and
colloidal
water are thus com-
pletely eliminated.
Fig. 133.
Apparatus for Scheibler's alcohol-extraction method.
Scheibler's Alcohol-extraction Method. The solvent most generused for the extraction of sugar from beet pulp is 90 per cent ethyl alcohol. The original method of Scheibler* as modified by Sickelf is
ally
as follows:
*
Neue Zeitschrift, 2, 1, 17, 287; 3, 242. t Ibid. 2, 692.
234
SUGAR ANALYSIS
normal
(or
double normal) weight of finely prepared pulp is a weighing dish, 3 c.c. of lead subacetate (6 c.c. for weighed the double normal weight) are then added and thoroughly mixed with the pulp by means of a glass rod, adding at the same time 5 to 10 c.c.
rapidly in
The pulp is then transferred to the extraction of 90 per cent alcohol. Soxhlet of a extractor, of which Fig. 133 shows six in the cylinder form of a battery. The bottom of the extraction cylinder is covered
of felt or cotton; the pulp is washed in with 90 per wad cent alcohol, and pressed down so that its upper surface is below the upper bend of the siphon tube S. The top of the extraction vessel is
with a clean
then connected by means of a tight-fitting cork with the condensing tube C, and the bottom with the 100 c.c. flask F, which should contain about 75 c.c. of 90 per cent alcohol. The water in the bath is heated until the alcohol in the flask begins
to boil vigorously,
ature.
side
when the heat is regulated to this constant temperThe vapor from the boiling alcohol passes upward through the tube A and condensing in C drops back upon the pulp in B. As
soon as the level of alcohol in
rises
above the bend of the tube S,
the alcoholic solution of sugar siphons mechanically into the flask F. The distilling and siphoning are continued until all the sugar is extracted, which, according to the fineness of the pulp,
Immediately after usually requires from 1 to 2 hours. the last siphoning the flask F is disconnected, cooled to room temperature, the volume completed to 100 c.c.,
and the solution mixed, filtered, and polarized. A form of extraction vessel devised by Miiller
(Fig.
134) permits the withdrawal of a small sample of liquid from the siphon tube for determining the completion of
The opening at a is closed during operation with a stopper. To obtain the sample this stopper is removed, a few cubic centimeters of liquid are sucked up with a pipette and subjected to the a-naphthol test
extraction.
(page 341 ).
Fig. 134.
If
the test
is
positive, the stopper is replaced
Miil-
and the extraction continued

coloration
^n
-
until the reagent gives
no
tlon
determining sugar by the Scheibler process of let's extractor" extraction special care must be exercised to prevent of alcohol during filtration. The funnel should be covered evaporation with a watch glass and the filtrate received in a cylinder or flask with
Tsfxh"
narrow neck.
carded.
The
first
20 to 30
c.c.
of the runnings should
be
dis-
The
greater susceptibility
of
alcoholic
sugar solutions to
235
expansion and contraction with changes in heat and cold necessitates the maintenance of uniform temperature conditions during the polarization.
The
specific rotation of sucrose in ethyl alcohol is slightly

is
higher (0.1 degree to 0.2 degree) than in water; but the difference small that it falls within the limits of experimental error.
so
The method
of alcoholic extraction gives results considerably lower
than those calculated from the polarization of the expressed juice. The results of Scheibler previously quoted (page 230) show a difference of about 0.75 for the polarization of sugar beets.
Some authorities prefer adding the lead subacetate to the alcoholic This practice extract rather than to the pulp previous to extraction.
is attended, however, with some danger. One main object of adding the basic lead to the pulp is to neutralize any free acid which would otherwise invert some of the sucrose in the hot solution. In presence
amount owing
of alcohol, lead subacetate solution must be used in lowest possible to the danger of precipitating sucrose or of changing its rotation specific through formation of lead saccbarate.
method can be applied to the polarization other sugar-containing plant substances. With very dry materials the strength of the alcohol should be correspondingly reduced. With substances
alcoholic extraction
of fruits
The
and
all
is
containing reducing sugars in large amount, it desirable to omit the addition of lead subacetate, but
when
this is
done the substance
should be well mixed with powdered calcium carbonate to neutralize any free acid that might
cause inversion.
II.
Determination of Sugar in Plant Substances by Extraction with Water
Water
is
sometimes used instead of alcohol
in extracting sugar for the polarization of plant In such cases a process of persubstances.
colation
must be used
in place of distillation Fig. 135.
Section of
Zam-
owing to the danger of decomposition through

the prolonged boiling of aqueous extracts. As an example of the water extraction process the
aron's hot-water extraction apparatus.
Zamaron* method
for
determining sugar in sugar cane is given. The Zamaron extraction Zamaron's Water-extraction Apparatus. apparatus (Figs. 135 and 136) consists of a cylindrical copper vessel
*
"
Sidersky's
Manuel,"
p. 261.
236
SUGAR ANALYSIS
basket B of perV provided at the bottom with a small cock C. forated copper, provided with a tripod support, fits loosely within this copper vessel; 100 gms. of the finely divided pulp are transferred to
the basket, and 200
c.c. of
hot water poured
in,
the pulp being pressed
Fig. 136.
Battery of Zamaron's hot-water extractors.
down beneath the surface The contents of the vessel

the flame
off into
of the liquid by means of the plunger P. are then boiled for 10 minutes, after which
is turned down, the cock opened, and the hot solution drawn the 1000-c.c. graduated flask F, as much as possible of the The cock is then liquid being pressed out by means of the plunger. closed and the process repeated with a second portion of 150 c.c. water.
The
continued 6 times, making altogether about 950 to 975 c.c. After cooling and adding a few cubic centimeters of lead subacetate, the contents of the flask are made to 1000 c.c., shaken, The reading multiplied by filtered, and polarized in a 400-mm. tube.
process
is
of extract.
1.3 gives the polarization (degrees
Ventzke) of the sugar cane.
The
principal objection,
which has been brought against the Zam-
aron process, is the danger of incomplete extraction. Some idea of the probable magnitude of this error may be formed from the following
consideration
:

also that 6 extractions of the pulp are
237
Suppose a sugar cane to contain 18 per cent of sucrose; suppose
made and
that one- third of the
If the sugar is evenly diffused through all parts of the liquid at the end of each 10 minutes boiling, as is no doubt very nearly true, there would be the following percentages of sugar removed at each extraction.
liquid
is
retained
by the
fiber after
each extraction.
238
SUGAR ANALYSIS
not exist in the alcohol-extraction method, owing to the insolubility of dextrinoid substances in ethyl alcohol.
///.
Determination of Sugar in Sugar Beets by Methods of Digestion
The method of alcoholic extraction, although the most accurate and scientifically perfect, is not the best from a practical standpoint on account of the long period of time necessary for extraction, and also because of the rather fragile nature of the extraction apparatus. For the rapid determination of sucrose in sugar beets some one of the numerous digestion processes
digestion of the extraction
is
usually followed.
The
pulp is the complete diffusion of the sugar through the liquid, the solution f is made up to volume, allowing for the space occupied by insoluble matter, and then filtered and polarized.
digestion
method may be regarded in principle as a combination and juice-expression methods. A weighed amount of digested with 5 to 6 times its volume of alcohol or water. After
The first process of Rapp-Degener Alcohol-digestion Method. * employed alcohol, and is known as the Rapp-Degener method. The double normal weight of fine beet pulp is transferred to a 201.2-c.c. flask (the extra 1.2 c.c. being the estimated volume of the insoluble cellular matter in 52. gms. of pulp). The forms of flask shown in
Three to four c.c. of leadFig. 137 are convenient for the purpose. subacetate solution are mixed with the pulp and then about 150 c.c. of
90 per cent alcohol added. The flask is closed with a stopper containing a condensing tube
and placed
is
in a hot- water bath.
The
alcohol
gently boiled for 20 minutes, when diffusion of the sugar through the solution may be considered complete. The tube and stopper are rinsed into the flask and the volume completed
nearly to the
Fig. 137.
mark with 90 per cent

in
alcohol.
.
.
-Flasks
for alcoof
hohc .digestion
pul
The
-
flagk
,
ig
laced in the hot-water bath
beet
for 1 to 2 minutes, to secure
even mixing of
the contents and expulsion of air bubbles, and then allowed to cool slowly in the air for J hour. The liquid is then brought to room temperature and the volume completed to 201.2 c.c. with 90 per cent alcohol. The solution is then mixed, filtered and
200-mm. tube, using the necessary precautions to prevent evaporation and changes in temperature.
polarized in a
*
Z. Ver. Deut. Zuckerind., 32, 514, 786.

The employment
also
of alcohol in analytical
239
it
work
is
expensive;
was
found that with any coarse particles of pulp the diffusion of sugar through the alcohol was considerably retarded. Pellet* was accordingly induced in 1887 to devise a method for determining sugar in beets in which water was used for digesting instead of alcohol. The Pellet method may be carried out with either hot or cold water. Pellet's Cold- water-digestion Process. Twenty six gms. of finely
divided pulp are transferred by means of a jet of water into a 200.6-c.c. flask (the extra 0.6 c.c. being the estimated volume of the insoluble marc in 26 gms. of pulp); 5 to 6 c.c. of lead-subacetate solution are
then added and sufficient water to

mixing, the flask
is
fill the flask about two-thirds. After allowed to stand for 20 to 30 minutes to permit
Fig. 138.
"
is
Sans-Pareille
"
which
placed in the cell C, is forced in
press for preparing finely divided pulp. a semiliquid condition
The substance, by the piston P
through the fine openings at the bottom into a container underneath; the latter also receives any overflow of juice which escapes by the outlet T.
diffusion of sugar and allow enclosed air bubbles to escape. Water is then added nearly to the mark, any foam destroyed with a drop of The solution is well ether, and the volume completed to 200.6 c.c. the scale reading in a 400-mm. and tube; mixed, filtered, polarized gives without correction the polarization of the beet. With pulp of extreme fineness, such as is obtained with the "Sans-
Pareille" press (Fig. 138), the diffusion of sugar from pulp to water becomes almost instantaneous, and the solution can be completed to
volume as soon as
*
air bubbles have arisen. thus considerably lessened.
The time
of analysis
is
Deut. Zuckerind. (1888), 1229; (1889), 531.
240
Pellet's
SUGAR ANALYSIS
Hot- water-digestion Process.
If
apparatus
is
not avail-
able for obtaining pulp of suitable fineness, hot water should be used to promote the diffusion of sugar from the coarser particles of pulp.
Twenty-six grams of pulp, mixed with 6 c.c. of lead-subacetate solution, are washed into a 200.6 c.c. flask, water is added with shaking until the volume is almost up to the mark, and the flask heated in a boiling water bath for J to 1 hour, according to the fineness of the pulp. The
then immersed in cold water; as soon as the contents are of room temperature, the volume is completed to the mark. The remainder of the process follows as under cold-water digestion.
flask is
Kriiger,* in 1896, deKriiger's Cold- water-digestion Process. vised a water-digestion process, an interesting feature of which is that the use of normal weights and of volumetric flasks is entirely dis-
pensed with.
the following:
The
principle of the
method may be understood from

an average sugar beet of 5 per
The weight
of juice per 26 gms. in

is
24.7 gms. The specific gravity of the average beet juice is very nearly 1.07, so that the volume of juice in a normal weight (26 gms.) of pulp is 24.7 gms. -r- 1.07 = 23.08 c.c.
cent marc content
26
0.95
The amount
100
c.c.
is
of water necessary to complete this
volume
therefore 100
of juice to
23.08
76.92
c.c.
The
:
ratio of
normal
1
weight to
2.958
c.c., therefore, of water in the proportion of 3 c.c. to every 1 gm. of pulp yields a solution whose polarization in a 200-mm. tube will give the approximate
:
volume of added water is then 26 gms. 76.92 c.c. or in round numbers 1 gm. 3 c.c. The addition,
gm.
sugar content of the beet.
The automatic
and lead solution
is
pipette (Figs. 139, 140) for rapidly measuring water an essential feature of the Kriiger process. The
pipette is prepared in several sizes for approximate double-normal, normal, half-normal, and quarter-normal weights of pulp (i.e., approximately 50, 25, 12, and 6 gms.), the smaller sizes being used in polarizing
mother
beets,
where the quantities of pulp obtained by the Keil sam-
The pipette, which is fastened pler (p. 226) are small (8 to 14 gms.). to a fixed support S (Fig. 140), is provided at opposite ends with the
three-way cocks C and C', the movements of which are controlled by the double lever L. The lower inlet of the pipette is connected by the tube A to the vessel V which contains the "lead water" (9 vols. of water to 1 vol. of lead-subacetate solution). The upper outlet which
permits the escape of air is connected with the upright tube B. By raising L to the stop c (Fig. 139) the pipette is filled with "lead water,"
*
Deut. Zuckerind. (1896), 2434.
241
any overflow passing into the tube B. Upon dropping L to the stop d, the cocks are both. reversed, air entering through/, and the contents of the pipette being discharged through e into the metal weighing dish
D, which contains the weighed sample of pulp.
d
Fig. 139
Fig. 140
Kriiger's automatic pipette for sugar beet analysis.
The weight of pulp corresponding to each pipette is determined by calibration with water, as in the following example. The weight of distilled water discharged by a Kriiger pipette at 20 C. was found to
be 78.38 gms. The volume of the pipette in true cubic centimeters is then 78.38 -v- 0.9972 = 78.6 c.c. 78.6 ^ 3 = 26.2 gms., the weight of beet pulp corresponding to the pipette. " " After mixing the pulp and the weighing dish is lead water
covered and the contents allowed to remain for 20 to 30 minutes.
The
solution
is
then well
stirred, filtered,
and polarized
in
a 200-mm.
tube.
242
SUGAR ANALYSIS
Kriiger method, while not claiming extreme accuracy, is suffiOn account of its simexact for many purposes of analysis. ciently been method has the and widely used in such places as rapidity plicity
The
beet-seed nurseries, depots for purchase of beets, etc., where large numbers of samples have to be polarized with the least possible loss of time.
The occlusion of Sachs-Le Docte Process of Water Digestion. bubbles by pulp and the uncertainty of knowing whether such bubbles are completely absent before making up to volume have been
air
the principal objections against the original Pellet process of digestion. This error does not occur in the Kriiger method, where the volume of The necessity solution is established independent of any occluded air.
of employing irregular weights for each individual pipette and the use of insufficient water for the complete diffusion of the sugar during the
cold digestion have been raised on the other hand as objections against the Krtiger method. Sachs * and Le Docte f have met these difficulties by always taking the regular normal weight (26 gms.) of pulp for
and adding a constant volume (177 c.c.) of water and lead subacetate so that the final estimated volume of solution, regardless
analysis
of insoluble
marc or occluded air, is always 200 c.c. The constant-volume figure 177 c.c. in the Sachs-Le Docte process
is derived from the following consideration. Sachs assumes as the average marc and juice content of the sugar beet 4.75 per cent and 95.25 per cent respectively. For the normal weight (26 gms.) of pulp
there would then be 26 gms. X .9525 = 24.765 gms. juice. The average sugar content and density of juices from beets of different richness are given in the following table together with the calculated volume of
juice (24.765 -f- sp. gr.), the volume of lead-water solution (200 c.c. less the volume of juice) and the polarization error resulting from use of the constant volume 177 c.c.
TABLE
Sugar
in beet.
XLV

that by use of the constant volume 177 c.c. the calculated polarization error is too small to be detected
It is seen
243
upon the saccharimeter. The constant-volume pipette emis ployed in the Sachs-Le Docte process
shown
in Fig. 141.
three-way cock
K at
the bottom
serves for the inlet of
lead reagent and water at for the delivery of the 177
and
C and
mixed
c.c. of
The cap A at the solution through D. the overflow, is contop, which receives Instead of bottle. nected with a waste water and lead reagent drawing in the
" " solulead- water separately, a single used. be tion of proper dilution may
One
of the cock connections
may
thus
be dispensed with. By ing the capillary tube h upon
raising or lowerits
support
is
the capacity of the pipette at adjusted to exactly 177 c.c.
easily
The method
of operation is similar
to that in the Kriiger process. Weigh 26 gms. of pulp in one of the tared metal
beakers; the latter are of about 250-c.c. capacity and are provided with a tight-
cover of rubber; add 177 c.c. of water containing 5 to 6 c.c. of lead subficting
acetate solution (of about 30 Be.)
and a add shake thoroughly. drop Filter, of glacial acetic acid to the filtrate, and The scale polarize in a 400-mm. tube.
reading gives the polarization of the
Where many analyses have to be performed a large number of metal beakers are used, all of which are counterpoised against the same weight.
beet.
If the particles of pulp are coarse Sach8 . Le Docte autoFig 141i the Sachs-Le Docte process should be matj c p i p ette for sugar beet
carried
*
out by hot
digestion.*
The
analysis.
Sucrerie Beige, Oct. 15, 1908. Bull, assoc. chim. sucr. dist., 27, 180.
244 method
SUGAR ANALYSIS
of operation is similar to that just described, except that the metal beakers, after addition of the 177 c.c. of lead- water solution to the pulp, are each covered with a special pneumatic cap of rubber which prevents any loss by evaporation. Fig. 142 shows a water bath for
The metal beakers are the Sachs-Le Docte hot-digestion process. in minutes 30 a water bath After cooling for heated to 80 C. placed the beakers are well shaken, when the contents are filtered and polarized
in the usual
way.
Herzfeld* has slightly modified the Sachs-Le Docte process for hot The pulp is weighed into small copper cans, 11 cm. high, digestion.
Fig. 142.
Sachs-Le Docte bath for hot-water digestion.
mouth diameter. The cans are closed during digestion with rubber stoppers or with good corks covered with tinfoil. The blowing out of stoppers during digestion has been raised as an objection against the Herzfeld modification. Stanek and Urban f
recommend the use
gasket.J
of cans provided with a spring cap
6 cm. body diameter, and 4 cm.
and rubber
comparison of sugar determinations in beets by the Sachscold-
Le Docte
and hot-digestion methods and by the Kruger method
is
given in the following table.

terminations reported
*
The
results are the average of
many
de-
by
Herzfeld.*
t Z. Zuckerind.
t
Bohmen,
34, 625.
A very full description of methods for analyzing sugar beets and
a complete bib(Bull. 146,
liography of the subject from 1839 to 1907 has been compiled U. S. Bur. of Chem.)
by Bryan
245
246
SUGAR ANALYSIS
80 per cent alcohol and this solubility is diminished by the addition of The asparagine error is therefore negligible in the lead subacetate.
methods
of alcoholic extraction or digestion.
Variation in
Marc
Content.
Among
other sources of error peculiar
to the digestion methods
may be mentioned the difference in quantity and volume of insoluble cellular matter in the normal weight of pulp. This volume is in fact variously given by different authorities as 0.6 c.c.,* 0.75 c.c.,f 1.35 c.c.,{ and the digestion flasks have been correspondingly
graduated at 200.6
c.c.,
200.75
c.c.,
and 201.35
c.c.
Pellet has devised
a special digestion flask with 5 graduations at 200.0 c.c., 200.5 c.c., 200.75 c.c., 201.0 c.c., and 201.5 c.c., so that the chemist may vary the volume according to the weight and character of pulp. While the
volume most generally prescribed is 200.6 c.c. for 26 gms. of pulp, it is evident that this figure must be greater for wilted beets and less for unripe beets. In the same way the volume of lead-water solution in the Sachs-Le Docte process would be greater or less than 177 c.c. The polarization errors due to normal variations from the average of 4.75 per cent marc are considerably less than 0.1, but in extreme cases of wilted
or watery beets the alcoholic extraction method should be used as a
control.
The error due to imbibition or colloidal water (p. 229) has also been raised against the digestion methods. The average difference between the expression and extraction methods was found by Scheibler to be about 0.75 per cent, which difference represents the combined
influence of unequal composition of juice and of the colloidal water. In the digestion methods the 23 c.c. of juice is diluted to 200 c.c. or
nearly ninefold, so that the combined errors of the juice methods are reduced to less than 0.1. In the digestion methods the error due
to unequal composition of juice is largely eliminated; the residual error due to the so-called colloidal water must therefore be very
small.
tion
The agreement between the aqueous digestion and alcoholic methods upon normal sugar beets is usually very close.
extrac-
As
to
which of the water-digestion methods is preferable it may be said that if apparatus is available for securing pulp of extreme fineness the coldwater digestion is upon the whole less open to error. But for pulp of coarse or uneven character hot-water digestion should be used to insure
complete extraction.
*
"
Friihling's
Anleitung," 209.
t Fribourg's t Sidersky's
"
Analyse chimique," 253.
"Manuel," 241.

POLARIZATION OF PLANT SUBSTANCES CONTAINING BUT CENTAGES OF SUGAR
247
Low PER-
The methods previously described may be applied with minor modifications to the polarization of plant substances containing but
low percentages of sugar.
The
polarization of spent sugar-beet chips
and sugar-cane bagasse may serve as illustrations of the methods. Polarization of Spent Beet Chips by the Expression Method. While the water circulating through the diffusion battery removes most of the sugar from the beet chips, a small amount of sugar always remains unextracted; this residual sugar occurs for the most part
within the uncrushed
cells of
the beet.
It is necessary, therefore, in
squeezing out the water from diffusion chips to apply extreme pressure, in order to secure the maximum quantity of residual sugar. A polari-
amount
zation of the expressed diffusion water are sufficient for the calculation.
and a determination
of its
100 c.c. of the diffusion water pressed from a sample of spent Example. beet chips were clarified with 2 c.c. of lead-subacetate solution and the volume completed to 110 c.c. The filtered solution gave a polarization of 2.0 V. in a
400-mm. tube.
The water content
of the chips,
upon drying 10 gms.
at 100
to
is 2.0 X 1.1 2.2 V. Calling the sp. gr. of the waste diffusion water 1.000 (which can be done without serious = error) the polarization of a normal weight would be (26.00 X 2.2) -5- 100 0.572 V., or for a 200-mm. tube 0.29 V. The polarization of the spent chips would then be (90.5 0.29) ^ 100 = 0.26.
110 C. to constant weight, was 90.5 per cent. The polarization corrected for the dilution
Polarization of Dried Beet Chips
by the Alcoholic Digestion and
Dried sugar-beet chips have frequently undergone a change in composition through formation of water-soluble The aqueous optically active gums at the high temperature of drying. digestion method may then give a polarization different from the true
Extraction Method.
sucrose content.
In such cases
it is
recommended
to use the alcoholic
digestion and extraction method of Herzfeld.* half normal weight of the finely ground dry chips is digested in a hot-water bath with 50 to 60 c.c. of 60 per cent alcohol, adding
of the digestion flask are
3 to 5 c.c. of lead-subacetate solution, for 30 minutes. The contents then transferred by means of a little 60 per
cent alcohol to a Soxhlet extractor
and extracted under reduced

is
pres-
sure for 5 to 6 hours (see Fig. 143). The alcoholic extract up to 100 c.c., filtered, and polarized in a 400-mm. tube.
* Z. Ver.
then
made
248
SUGAR ANALYSIS
by Hot-water Extraction. Zamaron method of The may be employed upon for sugar cane. described manner as same in the Owing, howbagasse matter in of cellular amount much the to larger bagasse only ever, 50 gms. are taken for extraction. The extract is made up to 1000 c.c. and polarized in a 400-mm. tube. The reading multiplied
Polarization of Sugar-cane Bagasse
hot-water-extraction
by
2.6 gives the polarization of the bagasse. Extraction waters of very low sugar con-
tent
sometimes concentrated before Five hundred cubic centipolarization. meters of the neutralized solution are evaporated to somewhat less than the desired volume, and then made up to 100 c.c. or 250 c.c. for polarization. The saccharimeter reading is divided by 5 or 2 to obtain the
are
polarization of the extract. Polarization of Sugar-cane
Bagasse by
Hot-water
tion, in
Digestion. Bagasse is also the method of hot- water digespolarized by

case, however, it is necessary the percentage of fiber. The determination may be made by the methods
which
to
Fig.
know
143.-Her Zfeld's appaof the Hawaiian chemists.*

.
ratus for alcoholic extraction
under reduced pressure.
Determination of Fiber in Bagasse.
- One
strong linen bag,
The sample
and again
times.
after
is
of bagasse are placed in a out with an hydraulic press. pressed juice then treated with cold running water for two minutes,
hundred grams
and the
pressed, the
is
The bag
then placed in an
which the
fiber
dish for four hours at

is
two operations being repeated alternately five air bath at 125 C. for half an hour, is removed from the bag and dried in a shallow the same temperature. When an hydraulic press
not available, the sample
may
be treated in cold running water
for
12 hours and dried as above described.

Digestion of Bagasse. Fifty grams of bagasse are weighed in a tared flask; 500 c.c. of water containing 2 c.c. of 5 per cent sodium
carbonate are added, and the flask connected with a vertical condenser. The solution is boiled gently for one hour, the flask being shaken
thoroughly every 15 minutes. After cooling the flask is re weighed, and the weight of contents determined. The weight of contents multi* Hawaiian Planters' Record, 3, 317.

plied
249
by 2 gives the weight (W) of fiber and solution corresponding to 100 gms. of bagasse. Letting F = the per cent fiber in the bagasse, F = the weight of solution corresponding to 100 gms. of bagasse.
W
99
The aqueous
c.c.
extract obtained
by the hot
to 100
digestion
c.c.
of the solution are
made up
in
is squeezed out; with lead-subacetate
reagent, filtered,
and polarized
is
a 400-mm. tube.
The
polarization
100 jP
(P) corrected for dilution
.
,
and
this
reduced to a normal weight

for
26
of extract
is
-r
iuu
X -7^ yy
100
P=
26 P ~7^r yy
which value
a 200-mm. tube be-
comes
13 P QQ yy
The
polarization of the bagasse

.
is
then found by the
formula
13
(W- F)
OF
P(W - F)
-99-
POLARIZATION
SUBSTANCES
CONTAINING
INSOLUBLE
MINERAL
MATTER
The polarization of substances containing insoluble mineral matter can in general be carried out by the methods of extraction or digestion Certain classes of products, however, such as previously described.
soluble saccharates,
required.
carbonatation filter-press cake may contain sugar in the form of inand in such cases special methods of treatment are
As examples
of
methods to be employed several processes

cake
will
for the polarization of filter-press
be described.
If
Polarization of
saccharate-free
Filter-press Cake Free from Saccharate. press cake be triturated with a known quantity
of
water and the filtered extract polarized, the polarization of the cake may be calculated very closely, provided its moisture content has been determined.
II
50 gms. of press cake were ground in a mortar with 200 c.c. of solution (which should not be alkaline) was then clarified with a little dry lead subacetate and polarized in a 400-mm. tube. reading of 5.2 V. was obtained. The moisture content of the cake, determined by drying 10 gms.
Example.
water.
The
in a
hot-water bath to constant weight, was 45.6 per cent.
It
is
desired to
know the polarization of the cake. The weight of water in the 50 gms. of cake is 50 X 0.456 = 22.8 gms. The total volume of liquid (disregarding the slight increase in volume through solution of sugar) is then 200 + 22.8 = 222.8 c.c. The polarization of the
solution reduced to a
1.000,
normal weight of 26 gms. to 100 c.c. (calling the sp. gr. be done without serious error) is (5.2 X 26) -s- 100 = 1.35 V., which for a 200-mm. tube is 0.68 V., or 0.68 gms. of sucrose in 100 c.c. of
which
may
250
solution.
SUGAR ANALYSIS
This corrected to 222.8
c.c.
0.68
2.228
1.52,
the grams of
2 = 3.04, the polarization or percentage sucrose in 50 gms. of cake; 1.52 of sucrose in the cake, if no other optically active substances are present.
The above method of calculation is sufficiently exact for substances When the polarization is high, however, neglect low of polarization. of the increase in volume through solution of sugar and of the change In such cases the in specific gravity introduces a considerable error. method of extraction. determined some should be by polarization In sugar-house practice the determination of moisture in the press
cake is usually dispensed with, it being assumed that the volume of inThe normal weight of cake soluble matter in 26 gms. of cake is 4 c.c.
is
then
100 25). In give the polarization (104 26 50 of out and the volume cake are practice gms. generally weighed made up to 200 c.c.
will
:
: : :
cake, up to volume,
made up to 104 c.c.; or, if a when triturated, clarified with
100-c.c. flask
be used, 25 gms. of
lead solution,
and the
liquid
made
In the previous example

water to 200
5.2
c.c.,
if
the 50 gms. of cake had been

c.c.
made up with
volume
there would be 192.3
of solution (allowing 4 c.c. for
of insoluble matter in 26 gms.).

V., therefore 192.3
:
5.2
polarization for 222.8 c.c. of solution was 222.8 6.02, the calculated polarization of the
:
The
cake for a 400-mm. tube.
This for a 200-mm. tube would be 3.01, which
is
only 0.03 V. lower than the result previously found.
filter-press
Polarization of Filter-^press Cake Containing Saccharate. When cake contains insoluble saccharates, the sugar must be liberated from combination before the solution to be polarized is made
up to volume.
this result.
Several methods have been followed for accomplishing
Decomposition of Saccharate by
Means
of Acetic Acid.
The 50 gms.
heated
of press cake, after transferring with
water to a 200-c.c.
flask, are
to boiling, and acetic acid added drop by drop until all free alkali is neutralized. The solution is then cooled, clarified, made up to volume, filtered, and polarized as previously described.
method
The Decomposition of Saccharate by Means of Carbon Dioxide. is practically the same as that just described, except that a stream of carbon dioxide led into the solution is used for decomposing
the saccharate, instead of acetic acid. The frothing, caused by evolution of carbon dioxide, is the principal objection against the acetic-acid method, and the decomposition by
meansôf carbon dioxide usually requires considerable time. Methods have been devised, therefore, to decompose insoluble saccharates in other
ways.
One
of the
most common
of such
methods
is
the following:

The saccharates
251
Decomposition of Saccharate by Means of Ammonium Nitrate. of calcium are quickly decomposed by ammonium
nitrate with formation of free sugar, calcium nitrate, The reaction for monocalcium saccharate is
and ammonia.
Ci 2 H 22 OnCaO
Saccharate
+ 2 NH N0 + H
4 3
= dsHÔn + Ca(N0
Sucrose
3 )2
+ 2 NH OH.
4
In carrying out the process 50 gms. of press cake are ground up with 15 gms. of ammonium nitrate and 100 c.c. of cold distilled water. The mixture is then washed into a 200-c.c. flask, clarified with a little
lead-acetate solution,
made up
to volume,
and polarized
in the usual
way.
An
may
tion of free
objection against the ammonium-nitrate method is the liberaammonia, which in presence of the lead-clarifying agent
ammonia
precipitate a part of the sucrose as lead saccharate. in some cases causes a darkening of the solution;
The
free
contact
with the brass fittings of polariscope tubes may also color the ammoCare should be exercised, therefore, to prevent niacal solution blue. contact of the solution with copper or brass during the analysis.
In order to Decomposition of Saccharate by Means of Zinc Nitrate. eliminate the formation of free alkali Stanek* has proposed the em-
ployment of zinc nitrate

proceeds as follows:
for
decomposing the saccharate.
The
reaction
CisHÔnCaO + Zn(N0 3 ) 2 + H 2
Monosaccharate
Zinc nitrate
Ci 2
22
On + Ca(N0 3 ) 2
Calcium nitrate
Sucrose
+ Zinc Zn(OH)
hydroxide.
The
precipitated zinc hydroxide is removed with the insoluble mineral matter of the cake and a perfectly neutral filtrate is obtained. In carrying out the process a double normal weight (52 gms.) of press cake is thoroughly triturated with 100 c.c. of water; a few drops
of phenolphthalein indicator are then added, and a neutral solution of The volume zinc nitrate run in until the red color is just discharged. is then completed to 210 c.c. (10 c.c. being allowed for the volume of
insoluble cake
and zinc hydroxide), and the solution filtered and polarized.
the cane-
The methods, which have been described for polarizing products of and beet-sugar industry, may be applied equally well to the
maple and
products,
etc.
polarization of other sucrose-containing substances, such as
sorghum methods
preserves, confections, jellies, also be applied to the polarization of substances which contain other sugars than sucrose, the only change necessary to make
The same
may
Z. Zuckerind.
Bohmen,
34, 161.
252
SUGAR ANALYSIS
being in the constant for the normal weight. As an example of the application of saccharimetric methods to other sugars besides sucrose,
the determination of milk sugar in milk
is
selected.
SACCHARIMETRIC DETERMINATION OF LACTOSE
The normal weight of lactose for a sacchaVentzke rimeter with the sugar scale may be taken îs 32.9 gms. (see to the low percentage of lactose in milk (2 to 8 per Owing p. 197). the normal weight, and, as it is more double to is best employ cent) it convenient to measure the milk, tables have been prepared which give the volumes of milk corresponding to multiples of the normal weights The following table gives the volumes for different saccharimeters. of milk for 65.8 gms. which correspond to different specific gravities.
Polarization of Milk.*
TABLE XLVI
Giving the
Volumes of Milk Corresponding
to
a Lactose Double Normal Weight
Specific gravitv of
milk.
253
In carrying out the process, the volume of milk corresponding to the lactose double normal weight is measured into a 102.6-c.c. flask.
For
clarification either 1 c.c. of the acid mercuric nitrate, or
the mercuric- iodide solution

of 102.6
may
does no harm). The liquid is c.c., the extra 2.6 c.c. being the estimated volume of the preAfter mixing, the liquid is filtered cipitated casein, albumin, and fat.
in a 400-mm. tube; the scale reading divided by 4 gives the approximate percentage of lactose in the milk. * The volume of precipiWiley and Swell's Double-dilution Method.
30 c.c. of be used (an excess of either -reagent shaken and then made up to a volume
and polarized
and
tate in the preceding method varies according to the content of protein fat so that the fixed estimate of 2.6 c.c. is not always accurate.
For more exact purposes of analysis the double-dilution method of Wiley and Ewell may be used. The general principle of double dilution, due to Scheibler, has been considered on page 209. Two separate double lactose-normal-weight portions of milk are introduced into a 100-c.c. and 200-c.c. flask respectively. The same volume of clarifying agent is then added to each flask and the volume completed to the mark. The solutions are shaken, filtered, and read The reading of the 100-c.c. solution subtracted in a 400-mm. tube. from 4 times the reading of the 200-c.c. solution gives the reading corrected for
volume
of precipitate,
and
this reading divided
by 4
gives the
percentage of lactose in the milk.

Example.
by the above
flask.
The saccharimeter readings (400-mm. tube) of a milk analyzed method were 20.00 for the 100-c.c. flask and 9.80 for the 200-c.c.
19.20,
corrected for volume of precipitate is then (4 X 9.80) 20.00 and the percentage of lactose is 19.20-=- 4 = 4.80. The volume of precipitate according to the above observations would be
The reading
100 (2o.o19.2), 4e-e
(seep 210)
.
Leffman and Beam's Method.

protein are
clarification
When
volume
the percentages of fat and

of precipitate
known
in a milk, the
formed during
can be calculated according to Leffman and Beam f by the following method. Calling the specific gravity of milk fat 0.93 the volume of precipitated fat
is
found by multiplying the grams of
fat in the
weight of
sample by ^r-^
Analyst, 21, 182.
1.075.
"
t
In the same
way
the volume of the precipi"

(1896), p. 39.
Analysis of Milk and Milk Products
254
tated protein-mercury
SUGAR ANALYSIS
compound
is
found by multiplying the grams of

-
protein in the weight of sample

of fat
by -^= =
The sum
of the
volumes
and protein is the volume in cubic centimeters of the precipitate. For the polarization of evaporated or condensed milks the single
lactose-normal-weight of substance is taken. The method of analysis in other respects is the same as described for ordinary milk. The determination of lactose in milk by the saccharimeter is not
considered
upon the whole to be as accurate as by the gravimetric
method
of copper reduction.
considerable variation
is
frequently
found in the determinations by the two methods. In ten comparative determinations of lactose in condensed milk by different collaborators of the Association of Official Agricultural Chemists* an average variation of
0.30
was found between the
results
by the
optical
and by
the gravimetric method, the differences ranging from 0.03 to 0.90. In a series of comparative determinations by Patrick and Boylef upon
unsweetened condensed milks, the following results were obtained:

of precipitated fat
255
and proteids are
of course greater in case of con-
densed or evaporated milks. Polarization of Milk Sugar.

lactose
is
when
The optical method for determining to the easily applied analysis of commercial milk-sugar, other optically active compounds are absent. The lactoseis
normal-weight of sugar a little alumina cream;
made up to 100 c.c. with the addition of with dark-colored products containing milk sugar the solution of substance must be clarified, following the same methods and precautions as in the polarization of :*aw cane sugars. In polarizing milk sugar the saccharimeter reading must not be taken
until mutarotation has disappeared; the solution of sugar
is
either al-
lowed to remain in the tube until a constant reading is obtained or the mutarotation is destroyed by adding a few cubic centimeters of N/ 10 sodium carbonate solution at the time of making up to volume.
of simple polarization described in the present chapter be applied to the polarization of products containing may obviously and other sugars. But in practical work it is found glucose, maltose, that such sugars generally occur in mixtures with other carbohydrates, and the methods for their determination are accordingly given elsewhere.
The methods
INFLUENCE OF TEMPERATURE UPON SACCHARIMETRIC OBSERVATIONS*

Before concluding this chapter upon methods of simple polarization, the influence of changes in temperature upon the accuracy of saccharimetric observations should be considered.
It has been shown (p. 127) that with an increase in temperature the specific rotation of sucrose undergoes a decrease and the rotatory power of the quartz compensation an increase, the combined effect of
influences producing a decrease in the saccharimeter reading of a normal weight of pure sucrose of 0.03 V. for 1 C. increase in temperall
temperatures between 20 and 30 C. the general 0.0003 (t - 20) may be used for changing the V*\ 1 equation F Ventzke reading (V) of pure sucrose at any temperature t to the readature,
and that
20
for
ing at
20.
Saccharimeter Temperature Corrections.
a temperature correction, similar to the above, was
The employment of made by the
* For a full discussion of this question with bibliographic references see paper " by Browne, The Use of Temperature Corrections in the Polarization of Raw Sugars and Other Products upon Quartz Wedge Saccharimeters," read before Section V, Seventh International Congress of Applied Chem., London, 1909, also in J. Ind. and Eng. Chem. I, 567, and Z. Ver. Deut. Zuckerind., 69, 404.
256
SUGAR ANALYSIS
United States Treasury Department in 1897, in its polarization of the Treasury Department to sugars assessed for duty. The right of make such corrections in the observed saccharimeter readings was contested in the courts by several importers of sugar, who founded their case largely upon the claim that the rotation of pure sucrose is not appreciably affected by changes in temperature. The chemists representing the government were successful, however, in showing that the to specific rotation of sucrose is thus affected, and after a final appeal
the United States Supreme Court the case of the importers was dis-
missed for want of jurisdiction.*
The decision of the courts, which apparently justified the use of temperature corrections established for pure sucrose in correcting the polarization of all grades of raw sugars, has unfortunately seemed to many chemists sufficient authorization to use such corrections indiscriminately in the polarization of any and every kind of sugar-containing material.
tivities of
impure product
acids,
Since the saccharimetric reading of a raw sugar or other is simply an expression of the sum of the optical ac-
gums,
the various constituents, sucrose, glucose, fructose, organic etc., it is evident that a system of temperature corrections
which shall give the saccharimeter reading that would be obtained at 20 C., must correct for the variations produced by temperature in the specific rotation of all the optically active ingredients and not of the
sucrose alone.
Wiley's Temperature Correction Table. Wiley f has prepared a temperature table for correcting the readings of quartz wedge saccharimeters which is based upon the variations in the Ventzke scale
reading of normal and fractional normal weights of pure sucrose. This table has a range from 75 V. to 100 V. for temperatures between 4C. and 40 C.; the corrections are to be subtracted from
the observed readings,
ardization.
when the temperature of polarization is below and to be added when the temperature is above that of standUnited States
rections.
The method
Treasury Department Method of Temperature Corof temperature corrections devised by the
Office of
Weights and Measures of the United States Coast and Geodetic Survey and adopted by the United States Treasury Department for
use in the Custom-House laboratories, consists in increasing or diminishing the saccharimeter reading of each sugar solution by the variation
For testimony in this case see "Transcript of Record," U. S. Supreme Court, the American Sugar Refining Company, vs. The United States,
t J.
*
Am. Chem.
Soc., 21, 568.

in reading
257
which a standard quartz plate shows from the computed value of this plate for the temperature of observation. sugar The following report gives the temperature corrections in sugar
degrees for a quartz control plate tested by the United States Bureau Standards.
DEPARTMENT OF COMMERCE AND LABOR, BUREAU OF STANDARDS, WASHINGTON

ACCOMPANYING REPORT OF TEMPERATURE CORRECTIONS IN SUGAR DEGREES FOR QUARTZ CONTROL PLATE 233-B.S. 1910
Degrees
centigrade.
258
recting polarizations
SUGAR ANALYSIS
may be seen from the following diagram (Fig. correction for pure sucrose solutions, and the apthe which gives 144), for solutions of sugar-beet and sugar-cane prodcorrections proximate ucts (according to results obtained by Browne*), to be applied to the readings of the Ventzke scale for 1 C. increase in temperature. It will be seen that the correction for beet products is much nearer
+ U.U3U + 0.025
-1-0.020
|f0.015
g+ 0.010
P.+ 0.005
S n noo

TABLE XLVII
Showing
Effect of
259
Increase in Temperature
Products,
upon Browne
the Polarization of
f
Sugar-cane
No.
2 3 4 5 6 7 8 9
10
11
260
SUGAR ANALYSIS
TABLE XLVIII
Showing
Effect of
Increase in Temperature
Products,
upon Browne f
the Polarization of
Sugar-beet
261
counterbalanced, the result being that the polarization of these sugars This increase continues, as increases with elevation of temperature. the polarization diminishes (the percentage of fructose and other im-
20 for expurities being greater), until, at a polarization of about hausted cane molasses, an increase of 1 C. in temperature causes an increase of over 0.1 V. in the saccharimeter reading.
upon
Correction of Polarizations for the Combined Influence of Temperature Since the ingredient of the Rotation of Sucrose and Invert Sugar.
sugar products, whose polarization is most susceptible to the influence of temperature, is invert sugar, a more accurate method of correcting
saccharimeter readings is to combine the temperature coefficients of 0.0003 S (t - 20) sucrose and invert sugar as by the formula: P 20 = P - 0.0045 I 20) in which P is the polarization at t C., S the per(t
t
centage of sucrose and / the percentage of invert sugar. If the percentage of invert sugar is unknown the temperature correction for converting polarizations to 20
C.
may
be determined approxi-
mately by the following empirical equations:
For cane products, For beet products,
P = P + 0.0015 (P< P = P + 0.0006 (P 20

l
80)
50)
(t (t
20
- 20), - 20).
Such formulae as the above while more accurate than corrections which are based upon the temperature coefficients of pure sucrose, fail to give accurate results upon many individual products whose composition differs from that of the average type. Polarization at Constant Temperature. It is evident from the
foregoing that the method of applying temperature corrections established for pure sucrose to the polarization of sugar products in
Since it is impossible to devise a simple reliable temperature corrections that can be applied to the polarization of all kinds of substances, the one means of securing uniformity and accuracy in saccharimetric work is to make all polarizations at the temperature at which the instruments are standardized. Customhouse laboratories, arbitration laboratories, and all other laboratories, upon the results of which great interests are involved, should be equipped with cooling and warming apparatus for maintaining a uniform standard temperature throughout the year.
general
is
faulty.
method
of
The New York Sugar Trade Laboratory was the

ratory in the
first
testing labo-
United States to follow out the requirements of the International Commission for Uniform Methods of Sugar Analysis and make all polarizations at 20 C. The laboratory room and polarizing cabinet used for this purpose are insulated. In warm weather the air
262
is
SUGAR ANALYSIS
by an electric fan through ducts over cooling coils, fresh being introduced from outside according to the needs of ventilation. A small ammonia compressor driven by an electric motor serves for the work of refrigeration. The temperature can be controlled either
circulated
air
Fig. 145.
Refrigerating machine for constant temperature polarization
(New York Sugar Trade Laboratory)
automatically by means of a thermostat which operates dampers regulating the passage of air to and from the cooling box, or directly by
means
of the rheostats controlling the speed of compressor
lating fan.
Figs. 145
The
121.
general arrangement of the equipment
is
and ventishown in
and
CHAPTER X
METHODS OF INVERT OR DOUBLE POLARIZATION
THE methods of direct polarization, as previously explained, give percentage of sucrose only in the absence of other optically active subTo determine the percentage of sucrose when other optically stances.
active substances are present, the method of inversion or double polarization is used, the principle of which may be understood from the
following. Law of Inversion.
When a solution of sucrose is acted upon by some inverting agent, such as an acid or the enzyme invertase, the sucrose molecule is broken up or inverted, giving rise, by the addition of one molecule of water, to one molecule each of glucose and fructose, the mixture of these two sugars in equal amounts being termed invert
sugar.
This reaction,
known
as hydrolysis or inversion,
is
expressed by
the following equation:
C H22 O n
12
2
(18)
12
Sucrose (342)
Water
Glucose
(180)
Invert Sugar (360)
__
:
12
6.
Fructose (180)
It is seen
from the above that one part of sucrose
is
converted into
360
n
1.05263 parts of invert sugar.
Calling the specific rotation at
20 C. +66.5 for sucrose, and 20.00 for invert sugar (p. 174), the relation of the optical activity of one part sucrose before and after inversion will be 21.0526 or a de66.5 1.05263 (- 20.00) = 66.5
crease of 87.5526 in specific rotation.
This decrease for one degree of 87 5526 - = 1.3166. The the saccharimeter scale would therefore be genbo. o
eral
law of inversion* as applied to the determination of sucrose may then be stated as follows: The total decrease in the saccharimeter reading at 20 C. of the normal weight of product after inversion divided by 1.3166 gives the
percentage of sucrose when no other optically active ingredient is hydrolyzed and when the inverting agent produces no change in the specific rotation of the other optically active constituents present.
*
For a
fuller discussion of
the laws of inversion see page 659.
263
264
SUGAR ANALYSIS
invertase
this
is
The enzyme
fulfills
most perfectly the conditions above
named, and when
used as the inverting agent the percentage of sucrose in mixtures with glucose, fructose, invert sugar, maltose, milk sugar, etc., may be determined very closely by use of the factor 1.3166.
The
vertase, however,
most commonly used in optical analysis is not inbut hydrochloric acid, the presence of which, as shown on page 185, has a most pronounced influence in increasing the specific When hydrochloric acid is used for inverting, the rotation of fructose. factor 1.3166 must be modified according to the amount of acid used for
inverting agent
inverting, the concentration of the sugar solution, and the manner of conducting the inversion. The extreme susceptibility of fructose to
changes in specific rotation and composition makes it necessary in employing any method of inversion to adhere most rigidly to the rules
of procedure prescribed.
THE CLERGET METHOD OF INVERSION

The method
1849,
by
of inversion for determining sucrose Clerget,* who found that a solution of the
c.c.,
was devised
in
French normal
weight of pure sucrose in 100
reading
+ 100
degrees
upon the
saccharimeter, gave after inversion with hydrochloric acid a reading of 34 degrees at 20 C. The total difference 44 degrees at C. or
between the readings before and after inversion, correcting for the
fluence of temperature,
is
in-
expressed by the quantity
100t
(-44)-|
144-|,
being the temperature of the inverted solution at polarization. If P f ) between the direct represents the algebraic difference (P
polarization (P)
and the invert polarization
then the percentage (S) of sucrose by Clerget's formula

the equation
(P'} of a given product,

is
expressed
at 20
by
C.
If
the invert polarization
is
made
the equation becomes
S =
or ^-~oT'
The factor 1.34 is considerably
greater than the factor 1.3166 for pure aqueous solutions of invert sugar. Tuchschmidf who subjected the Clerget process to an exhaustive
analysis, arrived at the following formula,
100
D
Ann. chim. phys.
[3],
144.16035- 0.50578 1
26, 175.
Compt.
rend., 16, 1000; 22, 1138; 23, 256; 26, 240; t Z. Ver. Deut. Zuckerind., 20, 649.
265
The original Clerget formula does not differ sufficiently from this to warrant the greater labor of calculation involved in the use of the
long decimals. If the direct
and invert readings are made upon a polarimeter with
circular degrees the Clerget formula would be, for the weight (1 sugar scale = 0.34657 circular degrees),
German normal
100
D
-.50""
(1
100 D
49.906
sugar
.34657 (144
for
0.173
V
0.21719
circular
the
French
normal weight
100
scale
degrees),
D
.5
100
31.275
D
0.109
1
.21719 (144
One gram
of sucrose dissolved to 100 metric cubic centimeters gives
a direct reading of
15 249
ing of
^
of
1.333 circular degrees
and an invert readthe grams of su-
^Q~
=
c.c.
0.5865 circular degrees at
0C;
crose (C) in 100
any solution may be found from the polarimeter
reading before and after inversion by the equation
P-P'
(49.906- 0.173 Q 26
P-P'
1.9195
0.0067
The Clerget formulae, given above, are to be employed only when the following method of inversion prescribed by Clerget is followed. After. taking the direct polarization (p. 202), the clarified solution remaining
at 50
is filled
up
to the 50-c.c. graduation
mark
of a flask graduated
c.c.; concentrated hydrochloric acid is then added to the 55-c.c. mark, a thermometer is inserted, and the flask slowly warmed until the temperature reaches 68 C., 15 minutes being taken in the heat-
and 55
solution is then quickly cooled, filtered if necessary, and as polarized nearly as possible at the original temperature of making up to volume. The polariscope reading for a 200-mm. tube of solution must
ing.*
The
be increased by TV to correct for the dilution with acid. The reading of the inverted solution is sometimes made in a 220-mm. tube, when
no correction for dilution

*
is
needed.
The
is
there
also a slight loss
addition of the acid causes an elevation of 2 to 3 C. in temperature; from evaporation during the inversion. It is, therefore,
better to control the temperature by inserting the thermometer in a 50-55 c.c. flask filled with water and placed in the bath with the solutions undergoing inversion.
After cooling to
room temperature, the volumes are readjusted
to 55 c.c.
266
SUGAR ANALYSIS
In carrying out the inversion special attention must be paid to all If the temperature of 68 C., or the time of 15 minutes, is exa partial destruction of fructose may result; if the temperature ceeded, of 68 C. is not reached, or if the time of heating is less than 15 mindetails.
utes,
some of the sucrose may escape inversion. Care must also be taken to maintain a constant temperature in the polarization tube during the reading. Even a slight warming of the tube, as from handling, will affect the observation.
jacket for circulation of
A polarization tube provided with a water at the desired temperature is very de(See Fig. 111.)
sirable for polarizing inverted solutions.
Herzf eld's Modification of the Clerget Method. The original method of Clerget has been variously modified from time to time in
order to diminish the danger of destroying fructose and to secure better uniformity of conditions. The inversion method of Herzfeld,* which is
the one most generally employed at present,
is
as follows:
is
The
75 5
c.c.
normal weight (13.00 gms.) of product of water into a 100-c.c. flask; after solution
half
transferred with
of soluble matter,
is
of hydrochloric acid of sp. gr. 1.188 are added, a thermometer introduced and the flask placed in a water bath heated to between As soon as the thermometer in the flask indicates 72 and 73 C.
c.c.
69 C.
(2.5 to 5
minutes) the solution
is
kept at this temperature for
exactly 5 minutes, rotating the flask gently at frequent intervals to seThe entire time of heating, accordcure even distribution of the heat. ing to the length of the preliminary period, will vary thus from 7 J to 10 minutes, and should never exceed 10 minutes. When the 5-minute
heating at 69 C. is completed the flask is cooled as quickly as possible to 20 C., the thermometer is rinsed from adhering sugar solution and
the volume
made
all
to 100 c.c.
After mixing and
filtering,
the solution
is
the precautions previously mentioned. The polariis doubled to obtain the correct invert reading for a norscope reading mal weight of substance. The invert reading for 26 gms. of chemically pure sucrose under the
polarized with
above conditions
is
-42.66 V. at 0, or -32.66 V. at 20
is
Clerget formula, according to Herzfeld's modification, ~ _ 100 by the equation
C. The then expressed
142.66 -0.5*;
or,
if
the polarization be
made always
=
at 20 C.,
_
-
by
100
132^6
7 Qft ==07538Z>
* Z. Ver. Deut. Zuckerind. (1888), 38, 699.

Effect of Concentration
267
the Clerget Factor. The factor 132.66 in correct only for a solution containing the half normal weight of sugar to 100 c.c. For other concentrations than this the value of the invert reading will vary according to the general
on
the preceding equation
is
formula
P
f
/20
which
is
0.0676 c), or P' = 0.0676 c), in (31.78 (41.78 the invert reading upon the Ventzke scale and c the grams
= -
of sucrose in
100
c.c.
The
following table gives the value of the
factor 142.66 in the equation
S =
142.66-0.5*
for different concen-
trations of sucrose.
TABLE
XLIX
Giving Clerget Factors at Different Concentrations of Sucrose

for Herzfeld's modified Method
Grams
sucrose
in 100 c.c.
268
this
SUGAR ANALYSIS
same sirup inverted according to Herzfeld's method gave a reading of Substi20 (t) upon the negative scale, - 9.7 X 2 = - 19.4 (P').
9.7 (N) at
tuting these values in the formula,
we obtain
~ S the
100
141.84
[+60 -(-19.4)] +(0.05X9.7)- (0.5 X
_ ~
20)
amount
of sucrose present in the sirup.
If the direct and invert polarizations be made at 20 C. the Clerget and Herzfeld formulae become simplified as follows:
Clerget formula
100 (P
30
P')
144-2
(P 66
0.7463
(P-P');
Herzfeld formula
0.7538 (P
P').
The values
tures between 10
of the Herzfeld factor in the simplified formula for temperaand 40 9 C. are given in Table L.
TABLE L
Giving the Inversion Factors for Herzfeld's Modification of Clerget' s Method
at Different
Temperatures
Temperature.
269
Inversion at Ordinary Temperature. The dangers of too high or too prolonged heating in the Clerget determination may be avoided by The time necesinverting at the ordinary laboratory temperature.
sary to invert a half-normal weight (13 gms.) of sucrose in 100 c.c. of solution employing hydrochloric acid of 1.18 sp. gr. was found by Ham-
merschmidt
to be as follows
Temperature.
270
Effect of Fructose
SUGAR ANALYSIS
on
the Clerget Factor.
Owing
to the influence of
fructose a Clerget formula hydrochloric acid upon the polarization of sucrose based upon the inversion of pure by means of this acid is not
absolutely correct when applied to the analysis of impure products containing invert sugar, since the specific rotation of fructose is different in the neutral
and acid solutions before and

introduced, in fact,
if
after inversion.
the Clerget formula estabconsiderable error is in the examination of molasses, be lished for pure sucrose employed materials other containing considerable fructose. honey, jam, jelly, and
Effect of
Amino Compounds on
acid used for inversion ents than fructose.
may
The hydrochloric the Clerget Factor. also affect the polarization of other ingredi-
Low-grade molasses, plant extracts, and other
sugar-containing materials frequently contain considerable quantities of optically active ammo compounds such as asparagine, aspartic acid, glutaminic acid, leucine, isoleucine, etc., the optical activity of which
and acidity of the solution. This may be table which gives the approximate specific rotaseen from the following in alkaline solution, in water, and in derivatives tions of several amino
varies with the alkalinity
hydrochloric acid.
TABLE
LI.
[a\D.
Approximate Value for
271
It is evident Clerget Modifications for Impure Sugar Products. that to overcome the variations in specific rotation of fructose, amino compounds, etc., which occur in the presence and absence of hydro-
chloric acid, the original

fied in the case of
method
of Clerget
may be Clerget modifications which atgrouped into two general classes. tempt to equalize the conditions before and after inversion with hydrochloric acid. II. Clerget modifications which employ an inverting agent free from the objections of hydrochloric acid. Among the modifications of Class I may be mentioned the following. (1) Neutralizing the Free Acid after Inversion before Making the Invert Polarization. This modification is best carried out in the Herzfeld
in fact
for convenience
I.
method have
impure products. been devised and these
must be considerably modiSeveral such modifications of the
process of inversion. After cooling the solution the free hydrochloric acid is carefully neutralized by means of sodium hydroxide, using phenolphthalein as indicator, and avoiding any excess of alkali. After
neutralizing, the
volume
is
completed to 100
c.c.
at 20 C.
and the
in-
vert polarization
polarization
mends that
made in the usual way. In order that the direct may be made under similar conditions Saillard* recomsodium chloride, equivalent to the amount present after
neutralizing the hydrochloric acid, be added to a separate solution before making up to the 100 c.c. for the direct polarization. The fructose, amino compounds, etc., are thus polarized under similar conditions before and after inversion. The Clerget constant for this method is
determined by making a parallel analysis upon pure sucrose.

(2)
Making
the Direct Polarization in Presence of Hydrochloric
Acid
and Urea.
This modification, due to Andrlik and Stanek,f is based upon the retarding influence which urea (or betaine) exercises upon the inversion of sucrose with hydrochloric acid in the cold. Fifty cubic
centimeters of the solution for the direct polarization are made up to 100 c.c. with a solution containing 5 gms. urea and 5 c.c. strong hydrochloric acid per 50 c.c. of reagent. After mixing, the solution is filtered
and polarized as quickly as
possible.
It is
claimed by the authors of the
method that a
sion
is
may
minutes) elapses before invernoticeable to make the direct polarization. While this claim be true for certain classes of products, it is certainly not the case
sufficient interval (7 to 10
with substances rich in sucrose.

presence of 5
*
The
comparison of the rate of inversion of 13

c.c.
following experiment shows a gms. of sucrose at 20 C. in

c.c.
strong hydrochloric acid and in presence of 5 acid hydrochloric plus 5 gms. urea in 100 c.c. of solution.
Eighth
Int.
strong
Cong. Applied Chem., Communications Vol.
XXV,
p. 541.
t Z. Zuckerind.
Bohmen,
31, 417.
272
SUGAR ANALYSIS
TABLE LII
Showing Influence of Urea upon
the
Rate of Inversion of Sucrose

It is seen that the 5
273
different rotation
gms. urea + 5 c.c. hydrochloric acid produce a than the 5 c.c. hydrochloric acid alone, this difference
This explains the high levorotation of of glucose ureide ([O\D = 23.5). invert sugar solutions prepared in presence of urea. (See Table LII.)
being greater for fructose. On long standing, glucose in presence of hydrochloric acid and urea shows a loss in rotation owing to the formation
The Andrlik-Stanek method is a dangerous one for it may introduce greater errors than those which it was designed to correct. The process, notwithstanding several favorable notices in the literature, is not to be generally recommended. Among the modified methods belonging to Class II, which employ for the Clerget determination inverting agents less open to the objections of hydrochloric acid, may be mentioned the following:
Means of Organic Acids. A number of organic such as have no influence acids, especially pronounced upon the optical
(1)
Inversion by
in place of hydrochloric acid the Clerget method. Weber* showed by that in presence of acetic acid invert sugar had the same rotatory power Acetic acid, however, is an unsatisfactory reas in aqueous solution.
activity of fructose,
have been employed
for the determination of sucrose
agent for the Clerget determination on account of its very weak invertTolmanf has ing action (%%Q that of hydrochloric acid, see p. 663). tested the use of citric acid for the Clerget process and found that with
2 gms. of this acid to 100
c.c.
complete inversion of sucrose could be ac-
complished in 30 minutes at the temperature of boiling water. Under these conditions the Clerget factor] for the normal weight of sucrose
Tolman noted, for the half-normal weight 141.49. the inretarded the that of soluble acetates however, presence greatly citric the was of acid and that latter action consequently of no verting
was 141.95 and
value as an inverting agent with products which required previous with lead subacetate. This same objection would apply to other many organic acids. Another serious objection, as with hydroclarification
chloric acid, against the use of organic acids as inverting agents
is
the
difference in optical activity of contaminating amino compounds in the solutions used for direct and invert polarization asparagine, for in but in dextrorotatory solution, example, being levorotatory aqueous
presence of strong acetic acid. Oxalic acid | has also been
recommended
2 gms. of the acid being used for 100 c.c. of solution. * J. Am. Chem. Soc., 17, 321.
t Bull. 73,
}
as an inverting agent, This acid has
U.
S.
Kulisch. Z. ang. chem. (1897), 45.
274
a
is
SUGAR ANALYSIS
much
stronger inverting power than either acetic or citric acid, but the same objections previously stated. to open The employment of organic acids as inverting agents in the ex-
amination of impure sugar products has not been found upon the whole
to be satisfactory. (2) Inversion by
an inverting agent
dicated
by
The employment of yeast as Means of Invertase. in the Clerget determination of sucrose was first inKjeldahl* in 1881. O'Sullivan and Tompson,f in 1891, and
Ling and Baker { in 1898, extended the use of the method and more recently Ogilvie has applied it to the analysis of sugar-factory products. The yeast method of O'Sullivan and Tompson, as modified by Ogilvie,
is
as follows "
Four times the normal sugar weight
of the
sample are trans-
ferred to a standardized 200-c.c.
flask, defecated with the
minimum
amount
alumina cream added, then the liquid adjusted to bulk at standard temperature, well shaken, and filtered; 100 c.c. of the filtrate are measured by a standard pipette into a small beaker, sulphur dioxide passed in from a
of basic lead-acetate solution (sp. gr., 1.26), a little
siphon of the liquefied gas till a faint smell is perceptible (all the lead thus being indicated to be precipitated), then the liquid transferred to a 200-c.c. flask, made up to the mark, and well mixed. Now sufficient
calcium carbonate (dried) in fine powder to neutralize the excess of acidity, and a little recently ignited kieselguhr (to promote filtration)
are added, after which filtration follows. In this way a normal solution is obtained, which is sufficiently clarified to give a distinct polarimetric reading, is free from lead and excess of acidity, and is therefore
well suited for the invertase inversion. " Fifty cubic centimeters of the solution, prepared in the manner just described, contained in a 100-c.c. flask, are raised in a constant-
temperature bath to between 50 and 55 C., after which 0.5 gm. of washed brewery yeast and 2 drops of acetic acid are added and the temperature maintained as near 55 C. as possible for 4J to 5 hours. At the end of this time the liquid is cooled, and a little alumina cream or kieselguhr
added to
ture.
assist filtration,
and made up to bulk

in
then polarized jacketed tube at exactly 20.0 C."

clear filtrate
is
The
at standard temperaa lateral-branched water-
The Clerget factor determined by Ogilvie for the above process from experiments upon pure sucrose is 141.6.
*
t J. Soc.
Compt. rend. Lab. Carlsberg (1881), Chem. Ind., 17, 111.
1, 192.
f J-
Chem.
Int.
Sugar Jour.,
Soc. Trans., 59, 46. 13, 145.

from
"
275
Instead of employing yeast, a solution of invertase prepared theremay be used to advantage. Hudson* has developed a method upon this principle, which is described as follows:
in water, clarify
Dissolve 26 gms. of the substance to be analyzed for cane sugar with the usual substances (neutral or basic lead acetate
or alumina
Filter
cream or kaolin) and make up to 100 c.c. volume at 20 C. and read the polarization of the nitrate in a 200-mm. tube.
the excess of lead from the
filtrate,
if
Remove
lead has been used
as clarifying agent, with sodium carbonate or potassium oxalate, and To 50 c.c. of the filtrate add acetic acid by drops until the filter.
reaction
(p.
is
669),
acid to litmus, add 5 c.c. of the stock invertase solution and make up the volume to 100 c.c. Add a few drops of
toluene to the solution to prevent the growth of microorganisms, shaking so as to saturate, and allow to stand at any temperature between
20
sion
and 40 C. over
is
time
is
Under usual conditions about six hours' night. to accomplish complete hydrolysis." When the inverrequired the solution is read at 20 C. and the invert reading calfinished,
culated to the normal weight of substance. The Clerget factor for the above method as determined by Hudson from experiments upon pure
sucrose
is
141.7.
The
invertase
method
is
unquestionably the most ideally perfect of
the numerous Clerget modifications. No disturbances are produced in the specific rotations of fructose, amino acids, or other optically active
substances which
may accompany
sucrose
and no other substances
than sucrose are hydrolyzed except in the few special cases where raffinose, stachyose or gentianose may be present. The complications involved in the preparation of the invertase reagent, the uncertainty of knowing whether a given preparation is always of constant strength, and the long period of time frequently
necessary to accomplish inversion are the chief drawbacks against the use of the method in practical analytical work.
The
fully
inverting power of the stock invertase solution should be care-
determined from time to time by experiments upon pure sucrose and with any decrease in activity the quantity of reagent used for inversion must be correspondingly increased. The time of inversion can be shortened considerably by conducting the inversion at a temperature of about 55 C. To determine whether or not inversion is complete the closed flask or tube of solution may be warmed again to 55 C. for an hour and then, after cooling to 20 C., reread. If no change
in polarization is noted, the inversion
* J.
is
complete.
Ind. Eng, Chem., 2, 143.
276
SUGAR ANALYSIS
The invertase method will be found of especial value in research work and in controlling the results of other methods. In this connection, however, it should be noted that the influence of salts and
other impurities upon the rotation of the accompanying sugars introduces the same error as in other Clerget modifications.
CLARIFICATION OF SOLUTIONS FOR THE DETERMINATION OF SUCROSE
BY THE CLERGET METHOD

In the analysis of sucrose-containing products by the Clerget method, by means of basic lead compounds must precede and not This precaution is necessary, owing to follow the process of inversion.
clarification
the occlusion of a part of the invert sugar in the basic lead precipitate and the consequent diminution of the invert polarization. In so far as the work of analysis will permit, the solution for the direct polarization and that used for inversion should both be taken from the same clarified
filtrate after deleading.
The
following
method
of procedure is given as
an example.
about 100
of Deleading. Transfer 57.20 gins, of product with of water to a graduated 200-c.c. flask. After solution, lead-subacetate reagent (1.26 sp. gr.) is added to the necessary point of
c.c.
Method
and the volume completed to 200 c.c. After mixing well, filtered and 100 c.c. of the filtrate (28.6 gms. substance) treated in a 110-c.c. flask with successive amounts of finely powdered potassium oxalate, or sodium carbonate, or sodium sulphate, etc., until no more lead is precipitated. If the deleaded solution is alkaline to
clarification
the solution
is
litmus paper or phenolphthalein it is exactly neutralized with acetic acid and the volume completed to 110 c.c. The solution is mixed,
filtered,
and the
filtrate
(26 gms. substance to 100 c.c.) used for the
direct polarization.
after completing the
then inverted in a 100-c.c.
Fifty cubic centimeters of the same filtrate are flask, according to the method desired, and,
to 100
c.c.,
volume
polarized for the invert reading.

re-
The
by 2 gives the invert polarization. In this connection it should be remarked that with substances
latter multiplied
of basic lead for clarification the 5 c.c. of hydroquiring large chloric acid prescribed for the Clerget or Herzfeld inversion may be
insufficient
amounts
of the
on account of the formation of chlorides and the liberation weakly inverting acetic acid. In such cases it is usual to em-
ploy 6 c.c. of hydrochloric acid for making the inversion. Instead of the powdered salts above mentioned, concentrated sulphurous acid (prepared by saturating water with sulphur dioxide) has been
proposed by Pellet for deleading.
This reagent has certain advantages,

for, in
277
addition to precipitating excess of lead, it neutralizes any free alkalinity and at the same time acts as a bleach upon any coloring matter which might darken the solution for reading. The sulphur
added to excess for deleading, sufficient quanthe solution of tity (10 being taken to complete the volume from 100 to 110 c.c. This excess does no harm, as the acid in the cold is a
dioxide has even been
c.c.)
very weak inverting agent and has no immediate depressing influence upon the direct polarization. This excess of sulphurous acid has also the advantage of preventing the troublesome afterdarkening which frequently results from the inverting action of hydrochloric acid. Ogilvie*
and
is
claims as another advantage an equalizing effect in the conditions before after inversion in that both direct and invert polarizations are made
in acid solution.
It is evident, however, that the total quantity of acid not the same in both cases and that these different amounts of acid
will exercise a variable influence
upon the rotation
of fructose,
amino
compounds,
etc.
An objection against sulphur dioxide as a deleading agent is the very troublesome character of the lead-sulphite precipitate which, on account of
the
its finely
filter.
divided colloidal condition, is very apt to pass through Agitating the solution with paper pulp, infusorial earth
(kieselguhr), or kaolin previous to filtration has been recommended as a means of securing a clear filtrate. Decolorization of Inverted Solutions. The afterdarkening which results from the action of the hydrochloric acid upon coloring sub-
stances, caramel, or other organic impurities, is frequently so great as to cause difficulty in reading the solution for the invert polarization.
In such cases a number of expedients may be followed. Since shortening the (1) Use of a 100-wra. or 50-mm. Tube. length of the observation tube always necessitates a corresponding multiplication of any errors of observation this method is to be used
only as a last resort.
(2)
Decolorization by
Means
of
Bone Black.
Animal charcoal or
bone black should never be used upon solutions for direct polarization on account of its great absorptive power for sucrose. It may, however, be employed with comparative safety upon solutions of invert sugar, provided the char be previously purified by washing with dilute hydrochloric acid and water and then dried. Two to five grams (depending upon the coloration of the solution) of the finely ground bone black are placed in the apex of a folded filter and the solution to be treated poured through in successive portions of about 10 c.c. The
*
Int.
Sugar Jour. 13, 145.
278
first
SUGAR ANALYSIS
25 to 30
c.c. of filtrate
are discarded
and the remainder used
for
the invert polarization.
Zinc Dust, of Reducing Agents, large number of reducing agents have been used for decolorizing acid solutions of invert sugar. Zinc dust has been frequently employed for this purpose, the destruction of coloring
(3) Decolorization
by
Means
Sodium
Sulphite, Etc.
matter being due to the nascent hydrogen generated by the action of The powdered metal is added to the hydrochloric acid upon zinc.
the solution to be decolorized in successive small amounts, thus preventing a too violent evolution of gas with loss of solution.
Sodium sulphite and bisulphite have also been employed for decolIn this case the bleaching agent is orizing acid invert sugar solutions. the sulphur dioxide liberated by the action of the hydrochloric acid.
The use
employed.
of zinc
attended with serious danger, provided only the
and sodium sulphite as decolorizing agents is not minimum amounts be
General Reliability of the Clerget Method
reliable results
While the method of double, or invert, polarization gives perfectly upon pure sucrose, it is evident that the method has serious limitations when applied to the investigation of impure prod-
The influence of mineral and organic impurities upon the of sucrose and other sugars, and the lead-precipitate rotations specific The influence of error affect all modifications of the Clerget process.
ucts.
hydrochloric acid
upon the
specific rotations of
fructose
and
ammo
where
compounds
is
an additional source of error
in all modifications
Under the invert polarization is made in hydrochloric acid solution. such circumstances the chemist need not expect, under the most favorable conditions, to obtain upon products containing a mixture of sucrose with reducing sugars, salts, and organic impurities an accuracy much greater than 0.5 per cent; in certain cases the error may exceed 1 per
cent. The Clerget method gives therefore at best only an approximation, the degree of exactness depending not only upon the care and skill of the chemist, but also upon the nature of the substance being analyzed.
The introduction of excessive refinements in the method has usually proved a thankless labor and is not to be recommended. The employment, for example, of a Clerget factor elaborated to the fifth decimal (as in Tuchschmid's formula, p. 264) is of no possible value in practical work. In employing any of the numerous Clerget modifications it is
always advisable for the chemist to establish his own factor for the

particular conditions of the analysis.
279
This is best done by making a blank determination upon pure sucrose, or, better still, upon a mixture of pure sucrose with approximate amounts of the accompanying substances which are known to occur in the product undergoing examinaso doing the chemist will gain an idea of the reliability of his method, such as can be secured in no other way.
tion.
By
APPLICATION OF THE CLERGET
METHOD TO THE DETERMINATION OF SUGARS IN PRESENCE OF SUCROSE
rotation
sucrose occurs in presence of another sugar, whose specific not affected by the inverting agent, and no other optically active substances are present, the percentage (Z) of the accompanying
is
When
sugar
If
may
be determined as follows:
the direct polarization for the sucrose normal weight of substance, and S the percentage of sucrose by the Clerget method, then P S is the polarizing power of the accompanying sugar. The peris
centage Z may then be determined as upon page 200, by dividing the value 100 (P S) by the polarizing power of the accompanying sugar (Table XXXVI). The calculation may also be expressed in general
terms by the equation
_ 66.5
in
(P
S)
~wT
the specific rotation of sucrose and [ag that of the accompanying sugar. The method of calculation may be illustrated
which 66.5
is
by
several examples.
Example
polarization of
I.
-+
A
58.0
sirup
the percentages of
and an invert polarization sucrose and dextrose.
containing sucrose and dextrose gave a direct of 8.33 at 20 C. Required
Pe.ee.
Per cent dextrose
=
=
66 5
-
50)
10 per cent.
O-6.O
Example
II.
polarization of
A sirup containing sucrose and invert sugar gave a direct 21 at 20 C. Required 52 and an invert polarization of
z
the percentages of sucrose and invert sugar.
Per cent
_e =
=
:
(EO
(
cent.
Afi ^
Per cent invert sugar
î^
*
=10 per
cent.
2,0
280
Example III.
4.20 at 20
SUGAR ANALYSIS
A
sweetened condensed milk (26 gms. in 100 c.c.) gave a and after inversion in the cold a polarization of
direct polarization of -f 51.50
C.
Per cent sucrose
Required the percentages of sucrose and lactose. 100 [51.50- (-4.20)] = 5570 = -
Per cent lactose
66.5(51.50-41.99) s
o2.o
--
41.99 per cent.
. = 10n 12.05
per cent.
The percentages
no
of sugars calculated in this

clarified
greater degree of accuracy
manner have of course than the Clerget sucrose determination.

by means
of basic lead
With impure products
compounds there
may
be an appreciable error due to the occlusion of reducing sugars in

of
the lead precipitate.
Dubois for Determining Sucrose and Lactose in Milk Dubois* has applied the Clerget method to the determination of sucrose and lactose in milk chocolate. The usual procedure is somewhat modified in that 100 c.c. of water are added to the 26 gms.
Method
Chocolate.
of substance, a correction being afterwards applied for the increase in
volume through solution
of sugars. preliminary extraction of the chocolate with ether to remove fat secures a more rapid solution of The following method of solution may also be used. sugars. of water, cork
Transfer 26 gms. of the finely ground chocolate to a flask, add 100 c.c. and heat in a steam bath for 20 minutes, releasing the
Shake thoroughly pressure occasionally during the first 5 minutes. Cool to room twice during the heating so as to emulsify completely. mix and filter. add 10 c.c. of lead-subacetate solution, temperature,
After taking the direct polarization (a), delead the solution with dry potassium oxalate. Invert the deleaded solution according to Herzf eld's
(6), correcting for Calculate the approximate percentages of sucrose (S) and lactose (L) by the following formulae:
method and take the invert polarization
dilution.
(a
6)
110 ~
(o
X "
1.10)
-S
The approximate grams (G) of total sugar in the normal weight of chocolate are calculated from S and L, and the volume (X) of solution
= 110 estimated by the formula (G X 0.62), in which 0.62 is the The increase in volume caused by dissolving 1 gm. of sugar in water. corrected percentages of sucrose and lactose are then found as follows:
True per cent sucrose

*
=
U.
Q-rr
True per cent lactose

S.
^77)'
Or.
66,

The employment
of
281
is
an expansion
factor, as in the
above method,
permissible only in case of water-free substances and where no other inThe factor 0.62 is not absolutely gredients than sugars are dissolved.
correct for all concentrations, as
Sucrose dis-
is
seen from the following table:
282
or 0.14037
SUGAR ANALYSIS
gm. anhydride)
by
the hydrolysis
The calculation is 0.4876 V. method may then be expressed as follows
of raffinose
:
R=
in
P-P
0.4876
which R is the percentage of raffinose, P the polarization of the normal weight of product before hydrolysis and P' the polarization of this normal weight after hydrolysis.
APPLICATION OF THE INVERSION
METHOD TO MIXTURES OF SUCROSE
AND RAFFINOSE
Raffinose is almost always associated in nature with sucrose, and since sucrose undergoes inversion simultaneously with the hydrolysis of raffinose, the formula previously given for the calculation of raffinose
Creydt,* however, showed that it was practical value. combine the equations for the calculation of raffinose and sucrose, and in this way obtain formulae which can serve for the estimation of the two sugars in mixtures. The original formulae of Creydt were based upon the old Clerget process of inversion and have now been largely replaced by formulae worked out for the Herzfeldf modi-
has but
little
possible to
fication (p. 266).
The method
of establishing these formulae
may
be
understood from the following:

If the sucrose normal weight (26.00 gms.) of a substance containing S per cent of sucrose and R per cent of raffinose (anhydride) be dissolved to 100 metric cubic centimeters and polarized in a 200-mm.
tube, the polarization of the sucrose in degrees Ventzke will be represented by S and the polarization of the raffinose by 1 .852 R (the value
nn
(\f\f\
1.852 being the ratio
of the
14.Uo7
normal weight
for raffinose
anhydride
to that for sucrose).
The
direct polarization
is
(the
sum
of the sucrose
and
raffinose polarizations)
represented then by the formula
P=S+
1.852 R,
whence
R=
^-^ l.ooz
and S =
P-
1.852/2.
(1)
If the sucrose normal weight of the above substance be inverted according to the Herzfeld method and polarized at 20 C., the invert polarization of the sucrose will be represented by -0.3266 S (since 1 V. sucrose before inversion reads 0.3266 V. at 20 C. after inversion).
In the same manner the polarization of the raffinose after hydrolysis will be 1.852 R x 0.5124 = 0.9490 R (since 1 V. raffinose before hydrolysis
reads +0.5124 V. at 20 C. after hydrolysis by Herzfeld's method).
*
t Ibid., 40, 194.

The
invert polarization
is
283
(the
sum
of the sucrose
and
raffinose invert
polarizations)
represented then by the formula
P'=- 0.3266 S + 0.9490 R.

By
substituting the quantity -pcHo"
(2)
(1) for
p_
o
f
equation
in equa-
tion (2),
we obtain the formula
whence
0.5124
P,
P'
0.839
Having calculated S from

r>
and
the value of
is
obtained from
equation
(1),
R=
substituting the quantity equation (2), we obtain the formula
By
1.852/2 of equation
(1) for
in
P'
= -0.3266
(P
1.852/2)
+ 0.9490 R,
.
whence
p _
0.3266
P + P'
1.554
By
direct
formula
(4)
the raffinose
may
be calculated at once, from the
and invert polarizations. The method of employing the formulae may be understood from
beet-molasses, free of reducing sugar, gave a direct polarization of 12 V. V. and an invert polarization of Required the percentages of
the following:
A
+50
sucrose and raffinose.
By
formula
(3),
per cent sucrose

per cent raffinose
= =
5124
*
'
^~
=
50
~ 12)
44.84 per cent.
By formula (1),
50
44 84 L.OO&
2.79 per cent, or
By formula (4), per cent raffinose =
0-3266
(-
12)
I.OO4
3.79 per cent.
Correction of Raffinose Formula for Changes in Temperature. The determinations of sucrose and raffinose by the preceding formulae
must be carried out at exactly 20 C.

other
In case the analysis is made at Several the formulae require to be modified. temperatures formulae have been worked out for correcting the invert polarizations of sucrose and raffinose for changes in temperature. Among the
simplest of these are the formulae of Herles,* which are derived as

follows:
*
Z. Zuckerind.
Bohmen,
13, 559; 16, 528.
284
SUGAR ANALYSIS
26.000 gms. of sucrose and 14.037 gms. of raffinose anhydride which read 100 per cent upon the saccharimeter before inversion, give after inverting by Herzf eld's method the following:
Temperature.
285
Reinhardt's explasorption of the highly dextrorotatory melibiose. nation is no doubt correct as bone black shows a similar absorptive
power
for other disaccharides, such as sucrose. Davoll,* who has made a detailed study of methods for estimating raffinose, gives the following results upon a mixture containing 94.98 per cent pure cane sugar
and 5.02 per cent
raffinose
hydrate (4.26 per cent raffinose anhydride).
The direct polarization for a normal weight of this mixture was +102.48. The invert polarizations for different methods of treatment were as
follows
:
Method
of treatment.
286
employed, as
in
is
SUGAR ANALYSIS
sometimes done, as a test for the presence of
of sucrose the chemist
raffinose
unknown mixtures. As in the Clerget determination
need not
expect in the analysis of commercial products for raffinose an accuracy much exceeding 0.5 per cent. The indication of a smaller amount of
raffinose
ities
than 0.5 per cent
is,
in fact, not regarded
by the
best author-
as sufficient to justify reporting its presence (as in raw beet sugars). Before applying the method to the analysis of unknown products
first satisfy
the chemist should
himself of the presence of raffinose
by
suitable tests (see p. 740); he should also confirm the results of his analysis so far as possible by making blank determinations upon known
mixtures.
A practical test of this kind

method
is
the best means for testing the
reliability of the
in particular cases.
CHAPTER XI
SPECIAL
METHODS OF SACCHARIMETRY
THE methods of inversion, described in the previous chapter, are only special instances of a more general course of procedure. It is possible to calculate the percentage of any sugar, provided its rotatory power, in distinction from that of associated sugars, can be given a
The changes definite alteration by some special method of treatment. produced in the rotation of sucrose and raffinose by the action of invertase or acids are but single illustrations of such special methods of treatment. As other examples may be mentioned (1) the determination of sugars by noting the change produced in polarization under different conditions of temperature. (2) The determination of sugars, by noting the change in polarization after fermenting with yeast.
(3)
The determination
of sugars
by noting the change
in polarization
after destroying the optical activity of reducing sugars.
Numerous
other examples might be given but the three cases cited are sufficient to illustrate the general application of the principle to special problems
of saccharimetry.
DETERMINATION OF SUGARS BY POLARIZATION AT HIGH TEMPERATURE

DETERMINATION OF INVERT SUGAR BY HIGH-TEMPERATURE
POLARIZATION
principle of this method is based upon the fact that solutions of pure invert sugar, when heated to a temperature between 85 and 90 C., become optically inactive. This inactivity is due to the lowering
in specific rotation of fructose
The
with increase in temperature (page 179)
the specific rotation of glucose being unaffected by temperature, the point of optical inactivity will be the degree at which the polarizing
powers of glucose and fructose exactly neutralize each other. The temperTemperature of Optical Inactivity of Invert Sugar. ature of optical inactivity of invert sugar has been variously estimated.
set the figure at
Dubrunfaut,* who made the earliest measurements of this constant, 90 C. Casamajorf and Wiley J have given 88 C.,
* t
Compt. rend., 42, 901. Chem. News, 44, 219. J. Am. Chem. Soc., 18,
287
81.
288
Lippmann,* 87.8 These variations
C.,
SUGAR ANALYSIS
may
Wolf,f 87.6 C. and Tuchschmid,t 87.2 C. be due in part to slight experimental errors
(such as incipient destruction of sugar at the high temperature) and in Inasmuch as the [O\D of glucose part to the influence of concentration.
cent solution to 54.0 for a 40 per cent evident that the temperatures at which these different polarizations are neutralized must vary somewhat. The effect of concentration upon the temperature of optical invaries
from
it
+ 52.5 for a 1 per
solution
is
activity for invert sugar may be determined established formulae of Gubbe.

||
by means
c.
of the carefully
Concentration
[a]
==
19.657
0.0361
II
Temperature
(20
20) In Table LIII, column B gives the [a] of invert sugar, as calculated by formula I, for different concentrations; column C gives the
[a]
(t
to 100 C.)
[aY D
+ 0.3246 - 20) - 0.00021
(t
grams
1
of invert sugar in 100 c.c. necessary to
produce a reading of
(page 197); column
1729
V., as calculated
by the expression 1UU
gives the temperature of optical inactivity, as determined by formula II of Gubbe; column gives the variation in degrees Ventzke, produced
by
gm.
is
of invert sugar in 100 c.c. for 1
and
calculated
by the expression n C (D
,
C. difference in temperature
p^20)
TABLE
A
LIII.
SPECIAL METHODS OF SACCHARIMETRY

The
is
289
easily understood.
application of the method to the determination of invert sugar Since a change of 1 C. produces a constant
variation of 0.018 V. for
1 gm. of invert sugar in 100 c.c., regardless of the concentration, then the grams of invert sugar in 100 c.c. of a given solution is found by the formula
Invert sugar
in
which and
P = P
f
Ventzke-scale reading at higher temperature t f Ventzke-scale reading at lower temperature t.
The method of applying the formula taking a typical example.
may
best be understood
by
50 gms. of a solution, containing a mixture of glucose and Example. fructose in unequal amounts, were made up to 100 c.c. at 20 C. The polarization was 10.20 V. at 20 C. in a 200-mm. tube.
50 gms. of the same solution were made up to 100 20.75 V. at 87 C. in a 200-mm. tube. polarization was
centage of sugars in the original solution.

Invert sugar
Q 7 P\
-^
-=
:
c.c.
at 87 C. The Required the per-
8.75 gn..
50
100
17.50 per cent invert sugar.
The
dextrorotation at 87
necessary to be paired with the fructose for invert sugar. may be estimated as follows
Since
1
C. shows an excess of glucose over the amount This excess of glucose
0.3225 gm. glucose (page 200) then the grams of glucose corresponding to the dextrorotation at the inactivity of invert sugar is 20.75 X 0.3225 = 6.69 gms. (uncorrected for concentration), or 13.38 per cent. To
scale
V.
correct for the influence of concentration, the true glucose value of the Ventzke0.0002 s 2 reading -f 20.75, according to the formula G = s -f 0.02 s
,
(page 199) is 21.08. 21.08 the original solution.
0.3225
6.80 gms. glucose or 13.60 per cent in
The percentage
of glucose
course, be considered as only approximate, for, as shown in temperature of optical inactivity, according to concentration,
determined by this method of calculation can, of Table LIII, the may be above or
below 87 C.
DETERMINATION OF COMMERCIAL GLUCOSE BY HIGH-TEMPERATURE

POLARIZATION
Method
of
Chandler and Ricketts.

first
The method
of high-tem-
developed in 1880 by Chandler and Ricketts * was not employed for determining invert sugar but for detecting the presence and estimating the amount of commercial glucose in cane
perature polarization as
*
J.
Am. Chem.
Soc., 2, 428.
290
SUGAR ANALYSIS
whose sugars, after inversion, sugar, molasses, honey and other products The material under examinaconsist almost wholly of invert sugar. tion was first inverted to convert any sucrose to invert sugar and then
polarized at the temperature of optical inactivity for invert sugar. dextrorotation observed at this temperature was attributed to
Any
com-
mercial glucose and

factor.
its
percentage estimated by means of an empirical
The
into
factor for converting the readings of the Ventzke sugar scale grams of commercial glucose depends entirely upon the nature of
Commercial glucose, as manufactured in the United in varies density from 41 Be. to 45 Be. (sp. gr. 1.388 to 1.442) States, 100 to 125 for the liquid and in specific rotation from about [a:]^
the product.
of commercial glucose corresponding to 1 product. of different products specific rotation are given in Table LIV.
The grams
V. for
TABLE LIV.
MD
(for
liquid product).

The use
291
of a special type of saccharimeter for high-temperature been largely discontinued. At present it is customary has polarization to make the polarizations upon an ordinary type of saccharimeter, employing a metal- jacketed tube; the latter may be insulated to advan-
tage
by a mantle
of asbestos or other non-conducting material.
The
FIG. 146.
Chandler and Ricketts's polariscope for high-temperature polarization.
hot water for heating the tube is conveyed by rubber tubing from a water-heater, which should be placed at a distance sufficient to prevent heating the polariscope. A convenient arrangement for this purpose,
described
by Leach,*
is
Method
by Leach. f
*
of Leach.
shown in Fig. 147. The following description
of a
method
etc., is
for
determining commercial glucose in molasses, sirups, honey,
given
"Food Inspection and Analysis"
t Bull. 81,
U.S. Bur. of Chem., p. 73.
(1911), p. 644. Bull. 107 (revised), U.S. Bur. of Chem.,
p. 74.
292
SUGAR ANALYSIS
"Invert a half-normal portion in the usual manner in a 100-c.c. flask; after inversion, cool, add a few drops of phenolphthalein and enough sodium hydroxide to neutralize; discharge the pink color with a
few drops of dilute hydrochloric acid, add from 5 to 10 c.c. of alumina cream, and make up to the mark and filter. Multiply by 2 the reading at 87 C. in the 200-mm. tube; multiply this result by 100 and
Fig. 147.
Apparatus for polarizing at high temperatures.
divide
by the factor 163 to express the commercial glucose in terms of 175 V." * glucose polarizing In the above method the solution is made up at room temperature
and polarized at 87 C.
for the
When this is done a correction must be made expansion of the solution and consequent lowering of the
The
best
test.
reading.
*
an empirical
method of making this correction is by means Thus Lythgoe,f following the above course
of the Association
of of
Provisional
Method
of Official Agricultural Chemists,
Bull. 107 (revised), U.S. Bur. of t Bull. 81, U.S. Bur. of
Chem., p. 71. Chem., p. 74.

procedure, obtained the following results
cial glucose.
293
upon
five
samples of commer-
Sample.
294
SUGAR ANALYSIS
TABLE LV
Kind
of honey.
295
cause a depression in the polarization difference, which will be proportional to the amount of commercial glucose used but irrespective of In order to correct for the variations in moisture its specific rotation.
and non-sugars
of pure
honey
it is
better to express the polarization
difference in terms of a uniform basis of 77 per cent reducing sugars, which is the average percentage of invert sugar after inversion for pure
honey.
The
formulae for
making the
100 (P
f
calculation are then:
Per cent pure honey =
P)
77
288.4 (P'
257
XI
^j
~V~
P)
Per cent commercial glucose = 100
in which P' = the Ventzke polarization of the inverted honey at 87 C. P = the Ventzke polarization of the inverted honey at 20 C. 7 = the per cent of invert sugar in the honey after inversion.
Another method, used in European countries, for estimating the of commercial glucose in honey is based upon the variation in the invert polarization of the sample from that of pure honey. Calling the average invert polarization of pure honey 17.5 at 20 C. (Table LV) and employing the official figure + 175 V. for the polarization of commercial glucose, then if
amount
= per cent of honey in sample, y = per cent of commercial glucose in sample, P = invert polarization of sample in degrees Ventzke, + y = 100.
x
0.175 x
y= -T93T
,
+ 1.75 y=P P + 17.5
This method of calculation, the same as that based upon the polarization at 87 C. makes no allowance for the wide range in the invert to polarization of individual honeys 15), so that a considerable
(30
error
may
be introduced in the
final result.
LVI the polarizations of 5 honeys and of mixtures of the with 20 same, per cent commercial glucose, are given together with the of commercial glucose as calculated by the three methods percentage
In Table
described.
be seen from the results in the table that with admixtures of low-purity honeys and commercial glucose there is a considerable error
It will
in the calculation of the percentage of added adulterant. The results obtained by any method for estimating commercial glucose have only an approximate value, and in no case ought such analytical results as
296
SUGAR ANALYSIS
those obtained for the pure basswood or white-oak honey to condemn a sample as being adulterated. In all suspicious or doubtful cases confirmatory qualitative tests such as that with iodine should be employed."
TABLE LVI
Polarization of
Honeys and Commercial Glucose Mixtures, with Calculated Percentages of Glucose by Different Formulce.

tions
297
of the
made
at
two widely separated temperatures by means
formula.
F=
in
P '~ P
0.0357 (Z'-O'
which
F = grams of fructose in 100 c.c. of solution. = Ventzke polarization at high temperature P = Ventzke polarization at low temperature
P'
factor 0.0357
t'.
t.
The
tions of other investigators as
employed by Wiley is confirmed by the observashown in Table LVII.

TABLE LVII.
Showing Change of Polarization of Fructose for
C.
Change of Temperature
298
SUGAR ANALYSIS
If 26 gms. of product are made up to 100 c.c. and polarized (P) at a low temperature t, and a second 26 gms. are made up to 100 c.c. and polarized (P') at a high temperature t', then the percentage of fructose F is determined by the equation
"
26
IQOCP'-P)
0.036
(t
-t)
_100(P -P) = 0.936 ('

t)
'
26 gms. of honey made up to 100 c.c. and polarized at 20 C. Example. 26 gms. of the same honey made up to 100 c.c. 14.8 V. gave a reading of and polarized at 87 C. gave a reading of -f- 10.50 V. Required the percentage of fructose.
In making polarizations at high temperatures it is desirable to make the readings as soon as the solution in the tube has reached temperature equilibrium, as indicated by the thermometer placed in the solution
and by the disappearance
of striations
from the
field.
After noting the
polarization the temperature is again taken and the average thermometer reading used in the calculation. Prolonged heating at high temA difficulty is sometimes peratures causes a destruction of fructose.
experienced in obtaining a clear unobscured field of vision when using the hot-water polariscope tube. Too slow a circulation of hot water through the jacket of the tube, with production of currents of unequally
heated solution, is the usual cause of the trouble. The hot water should be several degrees above the desired temperature and the circulation must be rapid enough to prevent loss of heat by radiation.
Limitations of
Methods
of
High-temperature Polarization.
The
by polarization at widely-separated temperatures, while giving good results upon dilute solutions of the pure sugars, gives only an approximation in case of many sugar mixtures. The method is strictly applicable only when the
of determining invert sugar or fructose
specific rotations of
method
in temperature;
in all other cases there will
the accompanying sugars are unaffected by changes be a certain error in the
determination depending upon the temperature coefficient and the percentage of other sugars present. While no other sugars are affected to the same extent as fructose, yet it must be remembered that 1.5 gms.
arabinose, or 3.0 gms. galactose, or 7.0 gms. maltose, or 9.0 gms. lactose, or 50 gms. sucrose produce approximately the same alteration in the
Ventzke reading with
C. variation in temperature as
this limitation the
gm.
of fructose,
or 2 gms. of invert sugar.
But notwithstanding
polarization has a distinctive value, and,
method of high-temperaturo when employed with due

caution, will be found of great service in and research.
299
of analysis
many problems
DETERMINATION OF SUGARS BY POLARIZATION BEFORE AND AFTER FERMENTATION
By employing pure cultures of specially selected organisms, it is sometimes possible to ferment one or more sugars of a given mixture, and from the variation in polarization thus produced to calculate the percentage of one or more of the members present. Action of Pure Yeast Cultures upon Different Sugars. The
fermentative action of various yeasts upon different sugars has been studied by Tollens and Stone,* Hansen,f Fischer and Thierf elder, J and
many
selective action
results of their experiments show a pronounced on the part of different yeasts. While pure cultures of such well-known yeasts, as Saccharomyces cerevisice, or Saccharomyces
others.
The
Pastorianus,
ferment
completely
d-glucose,
d-fructose,
d-mannose,
and maltose, these cultures are without action " milkupon 1-xylose, 1-arabinose, rhamnose, sorbose and lactose. A Fischer and Thierf fermented lactose sugar yeast," employed by elder, and sucrose completely but did not attack maltose. Saccharomyces apiculatus ferments d-glucose, d-mannose and d-fructose but not
d-galactose, sucrose,
galactose, sucrose, maltose or lactose. (See also Table CII, page 714.) Method of Fermentation. In carrying out experiments for the
by fermentation it is very essential that the culture be pure. The presence of foreign yeasts, moulds or bacteria may produce changes in sugars, which a pure culture would leave unattacked. The solution to be fermented should be sterilized
separation of sugars
of particular yeast
before inoculating. The most favorable conditions for the action of the yeast are obtained with a solution containing about 10 per cent sugar and kept at a tem-
perature of about 30 C.
It is also necessary, in order to secure a rapid and complete fermentation, to have a suitable supply of nutritive food matter present for the growth and sustenance of the yeast. supply for yeast in fermentation experiments is generally furnished by
means
of a nutritive salt solution or
by means
Hayduck's Nutritive Salt Solution.
of yeast extract. Dissolve 25 gms. potassium
phosphate, 8 gms. crystallized magnesium sulphate and 20 gms. asparagine in 1000 c.c. of spring water. One cubic centimeter of the above solution to each 25 c.c. of liquid
to be fermented insures a favorable
*
development of yeast.
J Ber., 27, 2031.
Ann., 249, 257.
t Centralblatt, 88, 1208, 1390.
300
SUGAR ANALYSIS
Wash 100 gms. of pure yeast (starch-free) reYeast Extract. peatedly with cold water and repress. The residue of yeast is then heated to boiling for one-fourth hour with 500 c.c. of water; the liquid is then filtered through a folded filter, the filtrate, in case of
turbidity, being returned to the filter until the extract runs through perThe extract is then made faintly acid with fectly clear. citric acid, when it is sterilized and preserved in flasks closed
by cotton wadding. The liquid to be fermented is diluted with an equal volume

of the
above extract. Fermentation experiments are best carried out in flasks closed with a washing tube for the escape of carbon dioxide. The apparatus shown in Fig. 148 answers very well for the purpose. The fermentation is continued until bubbles of gas cease to pass through the water in the washing tube, when the process is considered to be finished. The washing tube is then removed, the solution heated to expel all carbon dioxide, and, after cooling, clarified, and the volume com.
polarization of the filtered solution is calculated to unfermented sugar, and the difference in polarization, before and after fermentation, calculated to fermented sugar. The
tion flask.
The
application of the
method
is
best understood from a special case.
By hydrolyzing a sample of sawdust with sulphuric acid, the resultant liquid with an excess of powdered calcium carbonate, treating filtering and evaporating, a sirup resulted which contained the two sugars,
Example.
glucose
in a
and
xylose.
50 gms. of the sirup,
made up
to 100
c.c.,
gave a polarization of
+ 43.5
V.
200-mm. tube.
50 gms. of the sirup were then diluted in a 200-c.c. flask with 100 c.c. of water and 5 c.c. of nutritive salt solution. After sterilizing, cooling and inoculating with pure-yeast culture, the flask was closed with a washing tube and fermented for 5 days in an incubator at 30 C. The evolution of gas having ceased, the solution was heated to expel C0 2 cooled, clarified with a little normal acetate of lead solution, made up to 200 c.c., and filtered. The polariza,
tion of the filtrate in a

of glucose
400-mm. tube was
5.2
V.
Required the percentages
and xylose
in the sirup.
loss in polarization by fermenting was 43.5 - 5.2 = 38.3 V. Since 0.3225 gms. glucose in 100 c.c. then the grams of glucose fermented were 38.3 X 0.3225 = 12.35 gms. or 24.7 per cent glucose (unconnected) in the sirup.
The
V.
Since
1V.=
0.91
gms. xylose
in
100
c.c.,
then,
calling
the residual
SPECIAL
X
301
5.2 as due entirely to xylose, 5.2 0.91 = 4.73 gms. or 9.46 polarization of in the cent (uncorrected) xylose sirup. per Corrections for concentration are made as indicated on page 198.
Determination of Dextrin in Fruit Products. The fermentation is sometimes employed for the determination of dextrin in jams, jellies and other products, which might be adulterated with com-
method
The provisional method of the Association of Official * Agricultural Chemists is as follows " Dissolve 10 gms. of the sample in a 100-c.c. flask, add 20 mgs. of
mercial glucose.
:
potassium fluoride, and then about one-quarter of a cake of compressed Allow the fermentation to proceed below 25 C. for two or yeast. three hours to prevent excessive foaming, and then place in an incubator at a temperature of from 27 to 30 C. for five days. At the end
of that time, clarify with lead subacetate and alumina cream, make up to 100 c.c. and polarize in a 200-mm. tube. pure fruit jelly will show a rotation of not more than a few tenths of a degree either to the
If a polariscope having the Ventzke scale be used and a 10 per cent solution be polarized in a 200-mm. tube, the number of degrees read on the sugar scale of the instrument multiplied by
right or to the left.
0.875 will give the percentage of dextrin, or the following formula be used:
may
Percentage of dextrin
in
=
198
xLXW
which
C= L= W=
The
degrees of circular rotation. length of tube in decimeters. weight of sample in 1 cubic centimeter."
198 the [a] D of dexfactor 0.875 is found as follows: Calling are found from solution of then the of dextrin in 100 c.c. trin, grams (D) the Ventzke reading (7) in a 200-mm. tube by the formula:
gms. of product are made up to 100 c.c. then the percentage of 087^ V dextrin in the sample = X 100 = 0.875 V.
If 10
use of potassium fluoride in the method just described is to the prevent development of bacteria. Its employment is not necessary when pure-yeast cultures are used and the solution to be fermented has
The
been previously
*
sterilized.
Bull.,
107 (revised) U. S. Bur. of Chem., p. 80.
302
SUGAR ANALYSIS
of
The work
sice;
Brown and Morris * shows
that the dextrins and malto-
dextrins of starch conversion are not fermented
by Saccharomyces
cerevi-
their experiments prove, however, that other yeasts, such as Sac-
charomyces elUpsoideus and Saccharomyces Pastorianus, strongly ferment these dextrins. In carrying out the fermentation method for the estimation of dextrin, it is best to work with a pure culture of Saccharomyces
cerevisice.
Limitations of Fermentation Methods.
The methods
and
of estimat-
ing sugars by Several dangers attend the give at best only a fair approximation. employment of the method, chief among which are the attack of sugars, or carbohydrates, supposed to be unfermented, and the incomplete deCareful struction of sugars supposed to be completely fermented.
difference in polarization, before
after fermentation,
attention to the details of pure culture, sterilization and nutrition will, however, largely eliminate these dangers. The formation of optically active fermentation by-products may introduce a disturbing factor
under certain irregular conditions, but with a normal alcoholic fermentation the error from this cause is insignificant. The optical activity of the nutritive solution used in the experiments should of course be determined, and its value, if significant, should be considered in the
calculation.
The length of time required for completing a determination has been a strong objection against the use of fermentation methods in The more rapid, and generally more accurate, general sugar analysis. methods based upon polarizing and copper-reducing power have, for this reason, been given the preference.
POLARISCOPIC
METHODS BASED ON DESTROYING THE OPTICAL

ACTIVITY OF REDUCING SUGARS
The determination of sugars by methods of this class is based upon the fact that solutions of reducing sugars, when heated with alkalies or alkalies and hydrogen peroxide, or with alkalies and metallic oxides or
salts, lose
more or less completely their optical activity. These methods have been applied not so much to the determination of reducing sugars themselves, as to the determination of sucrose, dextrin and
other non-reducing carbohydrates in presence of reducing sugars.
DESTRUCTION OF OPTICAL ACTIVITY OF REDUCING SUGARS BY MEANS OF ALKALIES Method of Dubrunfaut. The first efforts to establish a quantitative method in this direction were made by Dubrunfaut f in 1850.
*
J.
Chem.
Soc. Trans., 47, 527.
Compt.
rend., 32, 439.
303
Later investigators found, however, that the end-products in Dubrunmethod, obtained by the action of different alkalies upon reducing sugars, were not completely inactive, so that the polariscopic reading Efforts to establish a constant always required a certain correction.
faut's
correction factor for modifications of Dubrunfaut's
method have been
made by
but the
Pellet,* Jesser,|
results,
Koydl,} Bardach and Silberstein and others, on account of the variability in conditions, have not been
wholly satisfactory.
Method
of
Lobry de Bruyn and van Ekenstein.
The
rate of de-
struction of optical activity upon heating solutions of reducing sugars with dilute alkalies is illustrated by the following experiment taken from the work of Lobry de Bruyn and van Ekenstein; 20 gms. of
II
anhydrous glucose were heated with 10 c.c. of normal potassium hydroxide in 500 c.c. of solution at 63 C. The following decrease in
rotation
was noted:
Time.
304
SUGAR ANALYSIS
Recent experiments by Jolles * upon arabinose, invert sugar, lactose and maltose show that these
Method of Jolles.
glucose, fructose, sugars in 1 to 2 per cent solution are rendered optically inactive
by heat-
ing for 24 hours at 37 C. with T o normal sodium hydroxide while sucrose is completely unchanged by this treatment. Stronger solutions
than 2 per cent show usually a residual activity after the alkaline treatment; it is necessary, therefore, in Jolles's method to dilute solutions to 2 per cent reducing sugar before making the deterof reducing sugars
mination.
tion necessarily involves a considerable multiplication of
With substances containing much reducing sugar such diluany errors in

of
the polariscope reading.
have modified
follows
:
Bardach and Silbersteinj Bardach and Silberstein. method so as to include solutions of reducing sugar up to 5 per cent concentration. Their method of procedure is as
Method
Jolles's
c.c. of the neutralized sugar solution and make up to 50 c.c. with normal sodium hydroxide, thus making the solution TV normal alkaline. The solution is then polarized and a measured volume placed
Take 45
cm. high and 5 cm. diameter) and kept 20 hours at 36 to 39 by means of a thermostat, the beaker The solution is then cooled, made up to the uncovered. remaining
in a small
beaker
(8 to 10
C. for
The final polarization is corrected for original volume and repolarized. residual activity by means of an empirical factor, which in case of glucose was found to be as follows:
TABLE LVIII
Showing Change in Polarization of Glucose upon
Approximate
Warming
ivith
Dilute Alkali
SPECIAL
305
polarization must be increased by 0.25 to give the correct polarization equivalent of the residual sucrose, or other non-reducing carbohydrate
present.
It is evident that the chemist in employing such methods as the above must establish his own correction factor for the particular reducing sugar with which he is working. The lack of absolute uni-
formity of conditions in the analysis of impure sugar products, leaves the general reliability of such correction factors more or less in doubt.
DESTRUCTION OF OPTICAL ACTIVITY OF REDUCING SUGARS BY MEANS OF ALKALI AND HYDROGEN PEROXIDE
Other chemicals have been used in connection with alkalies to pro-
mote the destruction of reducing sugars. Lemeland,* for example, has devised a method for destroying the optical activity of reducing sugars in presence of sucrose by means of alkali, manganese dioxide and hydrogen peroxide.
Method
which
is
of Pellet
recently proposed a
and Lemeland. Pellet and Lemeland f have method for the analysis of sugar-cane molasses,
by means
"
based upon destroying the optical activity of reducing sugars of alkali and hydrogen peroxide. The details of the method
a solution of the cane molasses that will contain at most
are as follows:
Make
5 per cent of reducing sugars.

300-c.c. flask,
Measure 50 c.c. of this solution into a sodium hydroxide (36 Be.), then 75 c.c. of hydrogen peroxide (12 vols.), and 60 c.c. of water. Mix and place the
add
7.5 c.c. of
flask in a boiling
maining alkalinity
water-bath for 20 minutes, cool, neutralize the refairly exactly with acetic acid, and defecate with
basic lead-acetate solution (36 Be.), the necessary amount of which will be found to vary from 15 to 40 c.c., according to the weight of the
material taken, the amount of reducing sugars destroyed and the imComplete the volume to purities initially contained in the liquid. First polarize directly in the 200-mm. or 300-c.c., mix well and filter.
400-mm. tube.
of glacial acetic acid
be taken, 1 c.c. added to it, the volume completed to 55 c.c., and after mixing a second polarization made, account being taken of the dilution. This is done because the second polarization is often a little If a difference different from the first, in which the liquid is alkaline. The is observed, then the second, or acid polarization, should be used. the on then and the on calculated sucrose is solution, percentage of
c.c.
Then 50
of the filtered liquid
may
sample."
* J.
t Int.
Pharm. Chim., Sugar J., 13,
2,
298.
616.
306
SUGAR ANALYSIS
state that the results
by this method agree very closely of inversion, when special premethod the with those obtained by utmost insure the to cautions are observed accuracy.
The authors
DESTRUCTION OF OPTICAL ACTIVITY OF REDUCING SUGARS BY MEANS OF ALKALI AND MERCURIC CYANIDE
The destruction of the optical activity of reMethod of Wiley. ducing sugars by means of Knapp's alkali-mercuric-cyanide solution was first employed by Wiley* in the determination of dextrin in comThe reagent is prepared as follows: Dissolve 120 gms. sodium hyAlkali-mercuric-cyanide Solution. droxide and 120 gms. mercuric cyanide in separate portions of water; the two solutions are then mixed and made up to 1000 c.c. Any
mercial glucose.
precipitate which forms
is
removed by
filtration.
In making the determination 10 gms. of the commercial glucose are dissolved in water and made up to 100 c.c.; 10 c.c. of this solution are
transferred to a 50-c.c.
graduated
flask,
20 to 25
c.c.
of the alkali-
mercuric-cyanide solution are added, and the mixture boiled 3 minutes under a well-ventilated hood. The solution is cooled, and neutralized
with concentrated hydrochloric acid, the latter being added until the brown color of the liquid is just discharged. The solution is then clarified,
made up
to volume, filtered
of the maltose
is
and polarized. The optical activity and dextrose being destroyed, the residual polarization
that of the dextrin.
taken as
In Wiley's experiments, the specific rotation of the dextrin was 193. Adopting this figure, and taking the reading of a Ventzke-scale saccharimeter, the grams of dextrin in 100 c.c. of solu-
tion
66 5
-
17O
tained
1
*.26 y0 =
=
Q Q896 yo
Since the golution p
ar i ze d con-
gm. of original sample
in 50 c.c. (or 2 gms. in 100 c.c.), then
0896
2
100
per cent dextrin in the commercial glucose.
In concluding this chapter upon special methods of saccharimetry the chemist is advised, as in case of the methods of inversion, to test the reliability of any untried process by means of check analyses upon mixtures of
known sugars. It is only in this way that an idea can be formed of the errors which are due to defect of method or to personal
*
equation.
Wiley's
"
Agricultural Analysis" (1897),
3,
290.
CHAPTER
XII
MISCELLANEOUS PHYSICAL METHODS AS APPLIED TO THE EXAMINATION OF SUGARS

IN addition to
tion there are a
lesser analytical
specific gravity, refractive
index and specific rota-
number
though of have nevertheless a considerable value in importance,
of other physical constants, which,
certain investigations of sugars and sugar solutions. Among the constants of this class may be mentioned viscosity, heat of combustion,
osmotic pressure, rate of diffusion, surface tension, heat of solution, thermal conductivity, specific heat and magnetic rotation. It is beyond the scope of the present volume to discuss the methods of making
each one of these physical measurements. Viscosity, heat of combusand the constants connected with osmotic pressure have acquired, however, a certain importance in general laboratory practice and the
tion
present chapter will discuss their use in the investigation of sugars.
VISCOSITY OF SUGAR SOLUTIONS
The determination
applied to
special
of viscosity of
solutions
is a measurement which is frequently and other carbohydrates for sugars
The purposes of technology, analysis or research. is an of a determined as arbitrary ordinarily viscosity liquid
is usually taken as the ratio between times of flow, a narrow tubular opening, of the same volumes of through water and liquid, all conditions of temperature, etc., being the same.
constant and
of
measurement
The simplest example of this method Viscosity Pipette. is afforded by the viscosity pipette. (Fig. 149.)
The pipette is first filled with water so that its meniscus coincides with the upper mark A; after holding in a perfectly upright position the water is released and the interval of time
noted for the passage of the meniscus from
to the lower
mark B.
Fl s- 149 -~ process repeated a number of times and the average result taken as the water constant of the pipette at The pipette is dried the temperature of the experiment.
The
is
and the process repeated

solution.
If
in exactly the same manner with a sugar the average time of flow at 20 C. for water be 20.2
307
308
SUGAR ANALYSIS
seconds and that of a sugar solution at 20 C. 105.1 seconds, then

105.1
20.2
5.2,
the relative viscosity of the sugar solution at 20
C, as
compared with water
of the same temperature. The apparatus of Engler* (Fig. 150) is Viscosimeter. Engler's used very generally for determining viscosity. The instrument conture.
sists of
The
a bath B, which is filled with water or oil of the desired temperacontainer A is gold plated, the conical bottom terminating
Fig. 150.
Engler's viscosimeter.
in a
narrow tube
the latter
a,
is
mm.
outlet;
closed
wide and 20 mm. long, which serves as the by the valve rod b. The container holds at
the marks
c exactly 240 c.c. of solution. After filling to c with water or solution, the cover A', holding a thermometer t, is placed in position
and the temperature brought to the desired point. The valve rod is then withdrawn and the time noted for the delivery of exactly 200 c.c. of liquid in the flask C. The calculation of viscosity is made as previously described.
*
Konig's "Untersuchung" (1898), p. 432.
MISCELLANEOUS PHYSICAL METHODS

Coefficient of Viscosity.
309
While the
viscosity, as calculated
it is
by
the above method,

in
is
sufficiently exact for
many purposes,
necessary
comparing liquids of different densities to employ the more exactly defined coefficient of viscosity. In Fig. 151 the volume V-oi liquid which is discharged in a time t through a given capillary tube A-B of the length I and radius r under a
pressure p
T7
is
found by the equation

TT
XpXr X SpXl
4
(1)
in
which p
is
the coefficient of the

It fol-
interior friction of the liquid,
lows from the foregoing that

_irpr
A
t
JL
(2)
Fig. 151.
Showing principle of viscosimeter.
When Vj r and I are unchanged, as happens in the use of the same viscosity apparatus, p under constant pressure p becomes
P
in
Kt,
(3)
which
is
a single constant peculiar to each individual viscosim-
eter.
In the previous figure the pressure

liquid
p,
with which a given volume of
discharged at the beginning of flow, is equal to its density d multiplied by the height h of its surface above the outlet B, and at the end of the flow to its density 8 multiplied by the height h'. In the disis
M-M'
M and M'
flow,
charge of a constant volume V of different liquids, between the marks h and h' are unchanged, so that for the mean pressure of
',
= C X
<5,
in
which C
is
a constant.
friction for different liquids using the
The coefficient of interior same viscosimeter is then repre-
sented by the formula

P
in
=KXCX8Xt,
For any liquid of density 8 and or ratio between the internal
which and For water (8

T,
C are two constants. = 1), p = K XC Xt.

is
time of flow
friction of
the viscosity coefficient
YJ,
water and liquid,
KXCX8Xr _8r KXCXt "V

The viscosity coefficients of liquids are, therefore, always proportional to the products of their densities and times of flow.
310
SUGAR ANALYSIS
The viscosity Viscosity Coefficients of Pure Sucrose Solutions. Orth* for differas determined sucrose of coefficients by solutions, pure
ent concentrations and temperatures, are given in Table LIX.
TABLE LIX
Viscosity Coefficients of
Pure Sucrose Solutions

For changes in temperature Orth gives the equation
311
or
in
log, (loge
t
ij)
loge (log.
A + log* B (x) + log C (t)

)
e
which x and solution, and A,
are the concentration
and temperature
of the sugar
and
constants.
300
250
200
150
Temperature
Fig. 152.
Diagram showing
viscosity curves of four sugar solutions
at different temperatures.
From the Viscosity Coefficients of Impure Sucrose Solutions. viscosity coefficients of solutions of different sugar-house products Orth has made a compilation, the results of which are shown in Table LX.
312
SUGAR ANALYSIS
TABLE
LX
Viscosity Coefficients of Sucrose Solutions of Different Purities.
Temperature.
313
Excessive viscosities may also occur in sugar-house practice from supersaturation of sucrose, the result of careless sugar boiling. The successful sugar boiler aims to prevent supersaturation and to keep the
mum
viscosity of the pan contents as low as possible, in order that the maxiyield of sugar crystals may be obtained. The determination of viscosity is of great value in certain branches
of analytical work, as, for example, the

trins, for
examination of commercial dex-
which see page 508.

SPECIFIC
HEAT OF COMBUSTION
The number of calories or when burned in oxygen under
Units Employed in Calorimetery. heat units which a substance gives off,

specified conditions,
is
investigation of sugars.
a constant which has been extensively used in the The determination has been especially emcalorific
ployed in studying the which are used in foods.
value of the different carbohydrates
The Small, or Gram, Calorie (cal.) is defined as the quantity of heat necessary to raise 1 gm. of water through 1 C. The quantity of heat to 1 C. is not, however, exnecessary to raise 1 gm. of water from
actly the same as that necessary to raise 1 gm. of water from 99 to 100 C., so that the measurement has been defined more precisely as one
one-hundredth of the heat required to raise

100 C.
gm. of water from
to
The Large, or Kilogram, Calorie (Cal.) contains 1000 small calories, and may be defined, with the limitations previously noted, as the
quantity of heat necessary to raise 1000 gms. of water through 1 C. The Centuple Calorie (K) is defined as the quantity of heat necessary
to raise 1
to 100 C. gm. of water from For ordinary purposes the ratio of the several units
1
may
be ex-
pressed as:
Cal
10
K=
1000
cal.
THE BOMB CALORIMETER

combustion is made in an atmosphere of compressed oxygen by means of a bomb calorimeter", the invention and extensive application of which to heat measurements are due to Berthelot.* The original bomb of Berthelot, on account of the
The determination
of calories of
amount of platinum which it contains, is exceedingly expensive, and has been variously modified by Mahler, Hempel, Atwater and
large
others for the purpose of reducing the cost.

*
The Berthelot
also
calorimeter,
[6] 6,
"Trait6 pratique de Calorimetrie chimique
"
;
Ann. chim. phys,,
546.
314
as modified
SUGAR ANALYSIS
by Hempel and Atwater* and improved by
Blakeslee,
is
shown
in Fig. 153.
calorimeter
The most important feature of the Description of Calorimeter. is the steel bomb, the cup (A) and cover (B) of which are lined with platinum, or heavily
plated with gold. The cover is provided with a sunken lead gasket K,
which rests upon the rim of the cup,
and
is
held in place
by the
steel
collar C,
which
is
screwed tightly
by means of a clamp and heavy spanner. The cover of

into position
is provided with a neck an having opening leading from G
the
bomb
to the interior of the
bomb
for the
is
entrance of oxygen;
the inlet
opened and closed by a valve screw F. The cover is also provided, on its inner surface, with two stiff platinum rods I and H, between which passes a small spiral of iron
wire for igniting the charge; the latter, consisting of 1 to 2 gms. of
the sugar or carbohydrate to be burned, is placed in a platinum

Fig. 153.
Bomb
calorimeter.
a small piece of act as a kindler, to naphthalene

capsule,
is
with
directly
under the
spiral.
The rod /
connected through the cover
with the electric wire /' and the rod H, insulated from the cover, with the electric wire H'
'.
The bomb, after introducing the Calorimeter. with charge, pure oxygen under 20 atmospheres pressure and then placed in the brittania-metal vessel M, which contains a weighed quantity of water, sufficient to cover all parts of the bomb. The vessel rests within two buckets, and 0, which, with their covers, form
Operation
of
is filled
two dead-air
spaces,
and insulate the bomb system from the room
at-
should be 2 to 3 C. mosphere. The temperature of the water in below that of the inner air-chamber. A Beckmann thermometer, P, passes through the covers of the pails, and is fastened so that its
*
See
article
by Atwater and
Snell, J.
Am. Chem.
Soc., 25, 659, for a
very com-
plete description of this instrument
and
its use.

bulb
is
315
immersed
in the
water about opposite the middle of the bomb.

read,
The thermometer can be
by means
of a magnifying
lens, to
the thousandth of a degree; it should be provided with a certificate for correcting errors of construction and for converting readings to The mercury thread of the thermometer is true centigrade degrees.
adjusted at the desired point by partly

reservoir.
filling
or emptying the upper

stirrer
When
in
the apparatus
is
in readiness the
mechanical
is
set
motion and the thermometer read at intervals of one minute, tapping the top gently with an electric hammer before each reading to prevent
When five successive readings show a lagging of the mercury thread. uniform rise in temperature, the electric switch is closed exactly at the
end of the
fifth
minute.
As soon
as the extinction of the
lamp
in a re-
sistance circuit indicates the fusion of the iron wire, the switch is reopened to avoid heating the water by the current. The readings of the
maximum
thermometer should be noted at the end of each minute, until the elevation of mercury is reached and the rate of fall has become regular. With the stirring mechanism making 40 revolutions per minute equilibrium is obtained usually within 5 minutes. After stirring 5 more minutes a final reading is taken, when the calculation may be made. The calories of combustion are calculated Hydrothermal Value. from the observations of a calorimeter experiment by multiplying the
hydrothermal value (in grams) of the calorimeter system by the corrected rise in temperature and dividing the product (after subtracting the heat units due to accessory combustions) by the weight in grams of
substance taken.
The accuracy
of all calorimetric experiments
is
dependent upon the
exactness with which the hydrothermal value of the calorimeter is known. The most common method for computing the water equivalent of the calorimeter system is to multiply the weight of each part by its
specific
heat and take the sum of these water equivalents as the hydrothermal value of the entire system. An example of the method is
given by Fries, in Table LXI. The hydrothermal value may also be determined by measuring the rise in temperature of the calorimeter system from burning a substance For a of known calorific value, as benzoic acid (1 gm. = 6322 cals.).
work
*
description of this of Fries.*

"
Fries,
and other methods reference should be made

in
to the
Methods and Standards

S.
Bomb
Galorimetry," Bull. 124, Bur. of
Animal
Ind.,
U.
Dept. of Agr.,
p. 9.
316
SUGAR ANALYSIS
TABLE LXI
Computed Water Value of Bomb Calorimeter
Material.

form for determinations of
Sample No.
of the experiment are given in the following record, this kind.
317
a convenient
which
is
Bomb No.
Description Cane Sugar. Date, July 13, 1901. Observer, J. F. Snell. Thermometer, No. 733.
318
SUGAR ANALYSIS
The weight of the iron wire was Correction for Accessory Combustions. 13 mgs. The quantity unburned was 1 .1 nig. The quantity burned was thereThe specific heat of combustion of iron being 1601 calories, the fore 11.9 mgs. 1.6 = 19 calories. The quantity of heat of combustion of 11.9 mgs. is 11.9
= 61.6 calories, the naphthalene burned was 6.4 mgs., which yields 6.4 X 9.63 The heat of specific heat of combustion of naphthalene being 9628 calories. combustion of nitrogen in the bomb as determined by titration of the nitric = 2 .004406 gm. HNO S = 1 acid is 6.6 calories. 2 3 5 (N 2
+H
HN0
cal.)
The total heat from accessory combustions is, therefore, 19
-f-
61.6
6.6 =
87.2 calories.
Deducting this quantity from the total heat set free in the apparatus, we 87.2 = 5403.6 calories as the heat due to the combustion of the have 5517.8 The quantity of sugar burned was 1.3718 gms. The specific heat of sugar. combustion according to this determination is, therefore, 5430.6 *- 1.3718 = 3959 calories.
of Combustion. The gram-molecular heat found by multiplying the calories per gram by the molecular weight (M ) To avoid large figures it is customary to express this unit in terms of large calories.
Gram-molecular Heat
is
.
of
combustion
Gm. mol.
Cals.
cals.
1000
CALORIFIC CONSTANTS OF DIFFERENT SUGARS
In Table LXII, compiled by Tollens,* the calorific constants are given for the principal sugars, polysaccharides and sugar alcohols. It is seen from the table that the molecular heat of combustion is
always higher for the anhydride than for the hydrate of the same sugar.
The molecular heat

greater than the
of
combustion of the higher saccharides
is
:
also
sum
of the values of their components.
Thus
"
Sucrose
=
675.9 J
1352.7
Gm.
mol. Cals.
Cals.
Glucose
Fructose
=673.7] = 1349.6 Gm. mol. = an* n r

3.1
Difference
Gm.
mol. Cals.
is liber-
This difference may be taken as the equivalent of heat which ated during inversion.
In the same way = 673.71 Glucose

Fructose
Galactose
*
Rafnnose
= =
2026.1
Gm.
Gm.
mol. Cals.
mol. Cals.
= =
675.9 V 669.9 J
2019.5
6.6
"
Difference
Tollens's
Gm.
mol. Cals.
"Handbuch
der Kohlenhydrate,
II, p. 45.

TABLE LXII.
Giving Heats of Combustion of Sugars, Poly saccharifies and Sugar Alcohols.
cal. 1
319
gram.
Cal.
for
1000 cal.) (1 Cal. 1 gram-molecule.
Sugars
Arabinose,
C H 10
5
Xylose,
10
6
Rhamnose, C H 12 O5
Rhamnose
Fucose, C H 12 O Glucose, C H 12 O Galactose, C H O Fructose, C H 12 O Sorbose, C H O Sucrose, Ci H 22 O u Lactose, Ci H 22 O n Lactose, Ci H 22 O u +H O

6 5
6
(cryst.),
CH O
6
12
+H O ....
2
3740 (IV 4379.3 (St. 3909.2 (St. 4340.9 (St.
3742.6 (St.)
3721 .5 (St.) 3755 (St.) 3714.5 (St. 3955.2 (St. 3951 .5 (St.
12
12
558.3 557.1 561.9 560.7 718.5 711.8 712.2 673.7 677.2 669.9 675.9 668.6 1352.7
1345.2 1340.6 1350.7 1339.8 1349.9 1345.3 2026.5 2026.1 2019.7 2043.0
(St.)
(B.)
(St.)
(B.)
(St.) (St.) (St.) (St.)
(B.)
(St.) (St.) (St.) (St.) (St.)
3736.8 (St.)
3949.3 (St. 3721 8 (St. 3947.0 (St. 3550.3 (St.
.
Maltose, Ci 2 22 O u 2O Maltose, Ci 2 22 O u Trehalose (anhydr.), Ci 2 22 O u Trehalose (cryst.), Ci 2 22 On+2H 2 O
H H
(Gibson)
(St.) (St.) (St.) (St.) (St.)
+H
H Raffinose (anhydr.), C H Oi Raffinose (cryst.), Ci H Oi +5 H O. Melezitose, Ci H Oi +H O

18
32
6
8
j
.
^'(B^'
3400.2 (St.) 3913.7 (St.)
(B.)
(St.) (St.)
32
32
Polysaccharifies:
Cellulose, (C 6
H O
10
5
5)
4185.4 (St.)
4182.5 (St.)
.
H O )n Dextran, (C H O )n.. Inulin, C 6H O Glycogen, (C H O )n

Starch, (C 6
10
6
10
62
3i
10
4112.3 (St.) 4133.5 (St.) 4190.6 (St.)
678.0 673.1 680.4 677.5 675.6 666.2 4092.1 678.9
(St.)
(Gottlieb)
(B.) (St.)
(Gibson)
(St.) (St.) (St.)
Sugar Alcohols:
Erythrite,
C H 10 O
4 6
4132.3 (St.) 4024.6 (St.)
CH O Mannite, C H H O6 Dulcite, C H O Perseite, C H O Quercite, C H O

Arabite,
12
5
3997.8 (St.)
3975.9 (St.) 3942.5 (St.)
4293.6 (St.)
.
14
16
12
Inosite,
CH O
6
12
3679.6 (St.)
504.1 (St.) 502 (Louguinine) 502.6 (B.) 612.0 (St.) 729.9 (St.) 720.5 (Gibson) 723.9 (St.) 836.1 (St.) 704. 4 (St.) 710.4 (B.) 662.3 (St.) 665.5 (St.)
St.
B.
= Stohmann and = Berthelot and
Langbein, J. prakt. chem. [2], 45, 305. coworkers, from results in the Ann. chim. phys.
[6], 6,
552; 10, 455;
13, 304,
341; 21, 409.
320
SUGAR ANALYSIS
hydrolysis of sugars
The
may
be regarded, therefore, as an exother-
mic reaction.
Various Calculation of Calories from Chemical Formulae. methods have been proposed for calculating the molecular heat of combustion from the chemical formula of sugars. The calorific value for the combustion of the elements carbon (diamond) and hydrogen have been determined as follows:
C + O = C0 + 94.3 Cals. H + O = H + 68.3 Cals.

2
Welter's * rule for computing the molecular heat of combustion is to form water from the subtract as much O and 2 as will unite to
molecular formula, and multiply the number of remaining atoms by The sum of the products is taken as the their respective heat values.
molecular heat of combustion.

Example.
to form 6
Glucose
Hi 2
H 0.
2
The
Cals. of the 6 remaining
This value
is
16 per cent less
of unite with 12 atoms of H atoms = 6 X 94.3 = 565.8 Cals. than the value found experimentally by Stoh6.
The 6 atoms
mann,
viz. 673.7 Cals.
the
A second method of calculating heat of combustion is to and C that will unite to form C0 and calculate the
2,
combine
all
heat of the
remaining atoms in the manner just described.
of
To take again the example of glucose The 6 atoms of to form 3 C0 2 The remaining C and Hi 2 then give
: .
unite with 3 atoms
For C, 3 For H 2 6
,
X X
94.3 68.3
= =
282.9 Cals.
409.8 Cals.
692.7 Cals.
The
results
by
this
method
are
much
closer
than those obtained by
Welter's rule, being about 3 per cent higher than the value found ex-
perimentally by Stohmann.
the
A third method of calculating heat of combustion is to distribute O of the molecule among its C and H atoms according to the pro-
Since the portionate number and combining powers of the latter. necessary to form C0 2 is represented by 2 C and the O to form 2 by
TT
>
-g
2
the uncombined equivalents of
C and
The
H, after deducting
CO
and
H O,
would equal 2
*
+ -^ - O.
TT
ratio of total to
uncombined
p. 129.
Walker's "Introduction to Physical Chemistry," (3rd Ed.),

equivalents
calculation
is
321
then ( 2
\
H + -= .
o)
/
-r-
(2
\
+ 5\
2>
The formula
for the
is
then:
-O
Gm.
mol. Cals.
=
^94.3
C + 68.3
we
Applying this formula to glucose,
obtain,
Gm.
mol. Cals.
(94.3 v
X 6 + 68.3 X
=
12
650.4,
'
+l
a result a
little
over 3 per cent below the value found experimentally by
Stohmann.
The
true molecular heat of combustion
is
about midway between the
values calculated
by the last two methods. It is evident, however, that absolute agreement cannot be attained by any method- of calculation, since the experimental results are different for different isomers.
The gram-molecule Calories Stohmann to vary from 668.6
for the
C Hi
6
sugars were found by
for sorbose to 675.9 for fructose.
OSMOTIC PRESSURE AND RELATED PHYSICAL CONSTANTS, AND THEIR APPLICATION IN DETERMINING MOLECULAR WEIGHTS OF SUGARS
The determination
derivatives
tion of
is
a problem which
of the molecular weights of sugars and sugar may confront the chemist in his examina-
unknown carbohydrates of plant or animal origin. In the case of a reducing sugar an elementary analysis of one of its osazones or hydrazones (p. 370) will serve to fix the class to which the sugar belongs and thus indicate the molecular weight. In the case,
however, of non-reducing sugars, such as sucrose, raffinose, etc., and of the sugar derivatives, which do not form osazones and hydrazones, a
determination of the molecular weight by some physical method
usually required.
is
The molecular weights of sugar derivatives, which can be distilled without decomposition or dissociation, are best determined by the wellknown vapor-density method of Victor Meyer. All the sugars, however, and most of their compounds undergo decomposition at or below
the melting point so that the vapor-density method is excluded. Recourse is, therefore, usually made to some one of the methods which in-
volve the principle of osmotic pressure.
322
SUGAR ANALYSIS
OSMOTIC PRESSURE OF SUGAR SOLUTIONS
Pfeffer,* the plant physiologist, in 1877,
upon osmosis
in vegetable cells, discovered that the
during his classical studies osmotic pressure of
Pfeffer's dilute sugar solutions was proportional to the concentration. experiments were performed by placing the sugar solutions in a porous bulb, which had deposited within its walls a semipermeable membrane
of copper ferrocyanide.
The
right tube,
was then immersed
bulb, which was connected with an upin distilled water. The membrane,
is permeable to water but not to sugar, allows water to enter the bulb; the sugar solution begins to rise in the tube and the elevation continues until, after many hours, a maximum is reached; at this point the difference between the level of liquids within and without the bulb
which
tion.
gives a pressure corresponding to the osmotic pressure of the sugar soluThis maximum pressure, expressed in centimeters or millimeters
was called by Pfeffer the osmotic pressure. following results by Pfeffer give the osmotic pressure of sucrose solutions at different concentrations.
of mercury,
The
Concentration

The
323
p
ratio
-^
is
thus also found to be constant, the slight variations
showed that
In 1887 van't Hoff* osmotic pressures were identical in value with those obtained by gas pressure; in other words that the osmotic pressure per gram-molecule of substance is the same as the gas pressure per gram molecule at the same temperature and volume. This identity is
Pfeffer's
being due as before to experimental errors. Relation of Osmotic to Gas Pressure.
expressed by the equation
pv
in
= RT,
which p is the pressure and v the volume, T the absolute temperature and R a constant. Van't Hoff showed that the constant R is the same
for substances in dilute solution as well as in the gaseous state.
The molecular weight

in
of a substance
is
equal to the weight of
its
grams which would occupy the same volume, under equal vapor temperature and pressure, as 2 grams of hydrogen (2 being the weight This volume, called the gram-molecular of the hydrogen molecule). c.c. at is C. 22,380 volume, (273 abs.) and 76.0 cm. of mercury pressure (1 atmosphere). Calling V the volume occupied from the previous equation,
by a gram-molecule
of gas
we obtain
*-*
pressure p, per square centimeter of mercury (sp. gr. = 13.59), is 13.59 = 1033 gms. We obtain, therefore, for the equal to 76 cm. constant R,
The
1033
22,380
~^73~
* 4 ' 683 '
To prove the identity of this constant for the osmotic pressure of sucrose one of the experiments of Pfeffer may be selected. A 1 per cent solution of sucrose at C. (273 abs.) gave an osmotic pressure of
49.3 cm. of mercury. The latter corresponds to a pressure per square Since the molecular weight of centimeter of 49.3 13.59 = 670 gms.
342, the volume (V) of a 1 per cent solution containing a gram-molecule would be very closely 34,200 c.c. Substituting these
sucrose
is
volumes in the equation, we obtain,
which value method.
is
in substantial
*
agreement with that derived by the other

"
Ostwald's
"
Grundriss
(2nd Ed.), p. 131.
324
SUGAR ANALYSIS
If we accept now the identity of the Application of the Method. and osmotic laws for gaseous pressure, the molecular weight of a sugar its osmotic from be determined can pressure in a manner analogous to
that followed
Example.
15.5
by the vapor-density method.

abs.) for a 1 per cent sucrose solution
C. (288.5
In one of the experiments previously cited Pfeffer found at an osmotic pressure of
52.05 cm. mercury. If 1 gm. of sucrose occupies 100 c.c. at 52.05 cm. pressure and 15.5 C., then C. (273 abs.) and the number of grams which would occupy 22,380 c.c. at 76 cm. pressure would be:
1
gm.
X
100
22,380
c.c.
c.c.
273
X 288.5 X 76 cm. = _ X 52.05 cm.
345 the number of grams in the gram-molecular volume is the molecular weight of sucrose. This agrees closely with the actual value 342 calculated from
the formula Ci2H 22 On.
It follows from the previous discussion that the sugars of lowest molecular weight will show for equal concentration and temperature the highest osmotic pressure.
Measurement
method
botanist de Vries,*
of
of
of applying the principle just described is
A second Osmotic Pressure by Plasmolysis. due to the Dutch

discovered that the plasmolysis, or loosening of cells, offered a simple and reliable means
who
the protoplasmic lining of plant
Fig. 154 shows the miscroscopic apIn of a cell in solutions of different concentration. plant sugar pearance such a cell the thin layer p of protoplasm (the protoplast) acts as a
measuring osmotic pressure.
semipermeable membrane.
I
So long as the osmotic pressure of the cell exceeds or that of the surrounding sugar solution s, the liquid equals is not affected. protoplast When, however, the osmotic pressure of
the sugar solution becomes greater than that of the cell liquid there is a The diffusion of water outward through the protoplasmic membrane.
latter, in
consequence of the
cell
loss of a part of the cell water, is loosened
from the
wall
and
contracts, as
shown
in the figure.
application of the method may be understood from the following: de Vries found that the hair roots of the frogbit (Hydrocharis Morsus-rance) showed no plasmolysis in a 7 per cent, but a very pro-
The
nounced loosening of the protoplast in a 7.1 per cent, sucrose solution. For these particular root hairs under the conditions of the experiment, plasmolysis was produced by a solution containing 0.208 gm. mol. of
sucrose to 1000 gms. of solution (71 gms. of sucrose).
*
-f-
342, the molecular weight
Bot. Ztg., 46, 229, 393.
325
Suppose that, using these same root hairs, a solution containing 3.7 per cent of glucose just produced plasmolysis. Then 37 (the grams of glucose per 1000 gms. of solution) divided by 0.208 = 178, the molecular weight of glucose, which corresponds to the formula C6 Hi2O 6 (molecular
weight =180).
Fig. 154.
I.
Condition of plant
Illustrating plasmolysis. before plasmolysis; II. Beginning of plasmolysis; III. Advanced stage of plasmolysis.
cell
It
was by
this
lecular weight of raffinose.
means that de Vries,* in 1888, established the moThe following formulae had been proposed
2
for the constitution of this sugar.

I.
II.
III.
+ 3 H = 396, molecular weight. = 594, molecular weight. H Ci 20 + 5 H = 1188, molecular weight. O 10H C36H +
Ci2H22 Oii
8 3
16
64
32
plasmolysis that, when standardized a sucrose solution for the same against plant cell, 595.7 parts of raffinose
De Vries found by his method of
were equimolecular with 342 parts of sucrose. This figure agrees with the molecular weight of formula II; the correctness of de Vries's conclusion was afterwards verified by chemical means.
Owing
that the particular plant cells chosen for this must always be standardized before using.
to the variation in composition of cell liquids, it is evident method of examination
FREEZING AND BOILING POINTS OF SUGAR SOLUTIONS
On
membrane and owing

of osmotic pressure,
account of the difficulty of preparing a perfect semipermeable to the extreme liability of such membranes to
rupture, the determination of molecular weights

followed.
by
direct
is
measurement
not generally
although most sound in principle, accordingly made of the measurement of some related constant, such as that of vapor pressure, depression of freezing * Compt. rend., 106, 751.
Use
is
326
SUGAR ANALYSIS
of point or elevation of boiling point. The freezing and boiling points sugar solutions vary in fact according to their vapor pressure, the value of which, it can be shown, is directly proportional to the osmotic
pressure.
In Fig. 155 suppose the closed vessel V to be M-M' into two equal compartments, which open into one another above M. Suppose, next, equal volumes of sucrose and glucose solutions
Isotonic Solutions.
divided
by a semipermeable membrane
same concentration to be placed Then compartments. water will diffuse from the sucrose soluof the in each of the
tion Sj where the osmotic pressure is lower, into the glucose solution G, where the osmotic pressure is higher, until at
of equilibrium the osmotic pressures upon both sides of the membrane are equal. The two sugar solu-
the
point
M'
tions are then said to be isotonic
and
Fig. 155.-Illustrating principle of isotonic sugar solutions.
isotonic solutions 'must have the same
vaP or pressure. For if the vapor pressures were unequal, water vapor would
pass from the solution of higher to that of lower vapor pressure, the concentration of the sugar solutions would thus be changed, and water must again diffuse to the compartment of higher osmotic pressure.
There would thus be established a perpetual motion which is contrary to law. Consequently isotonic solutions must have the same vapor pressure. Suppose next a piece of ice / to be placed in the closed compartment above the partition M, and suppose this ice to be of the same temperature as the freezing point of the isotonic sucrose solution S. Then the vapor pressure between 7 and S must be equal, otherwise water vapor would pass between the two and change the freezing point of S. But since S and G are both isotonic and have the same vapor pressure, both must also have the same freezing point. In the same way the two isotonic solutions S and G must have the
same
boiling point, the vapor tension of the aqueous vapor at the boiling point being the same for both solutions.
The proportionality between changes in vapor pressure and between changes in freezing or boiling point is easily illustrated by means of a be the pressure curve of water for diagram. In Fig. 156, let
OW
change in temperature and 01 the pressure curve of
ice,
the projection of
327
Let Ss be the corresponding at T being the freezing point of water. curve of a 1 per cent sucrose solution and Gg of a 1 per cent glucose f solution, the projection of the points s and g at t and t being the re-
For comparatively spective freezing points of the two solutions. r small areas the lines gO, ss' and gg may be regarded as straight and ss'
t'
T
Temperature
Fig. 156.
Showing
relation of vapor pressure of sugar solutions to depression in
freezing points.
and gg' as parallel. In the Os Og and so also Ogg', Os Og Tt Tt' Os Og Therefore the lowerings in vapor pressure (and hence osmotic pressure) Os' and Og' of the two sugar solutions as com: : :
: :
pared with the solvent water are directly proportional to the corresponding depressions in freezing point Tt and Tt' Raoult's Method for Determining Depression of Freezing Point. For determining the depression of freezing points by Raoult's * method the apparatus of Beckmann f (Fig. 157) is generally used. This con.
sists of
a large tube
(2.5
cm.
21 cm.) provided with a side tube A'.
The main opening is provided with a stopper through which pass the Beckmann thermometer D and a small stirrer, provided with a cork handle r. The thermometer has a range of about 6 degrees and the
scale
estimated
divided into hundredths, the thousandths of a degree being by aid of a magnifying glass. The tube A fits through a cork into the larger tube B, which serves as an air-jacket, and the
is
whole sets in the cover of a large glass cylinder which is filled with a freezing mixture a few degrees lower than the freezing point of the solution to be examined.
*
Compt.
t Z. physik.
rend., 94, 1517; 101, 1056; Chem., 2, 638.
103, 1125.
328
SUGAR ANALYSIS
In making an experiment, using water as the solvent, the freezing 5 C. and the mercury of the Beckmann therbath is set at about
mometer adjusted by means

its
of
regulating device c, so that the top of the column falls within
the proper range of the scale. weighed quantity of water, sufficient to cover the bulb of
the Beckmann thermometer, is placed in A, the thermometer and stirrer are inserted and the
tube plunged through the small opening b into the freezing mixture.
When
signs of freezing
begin to appear, the tube is withdrawn from the freezing

mixture, wiped dry and then inserted in the air-jacket B.
The water and forming ice now stirred vigorously by r;

temperature
certain
after
are
the a
reaching
begins to increase suddenly with the libThe eration of latent .heat.
minimum
mercury soon ceases to rise and the point at which it stops, after
tapping to prevent any lag, is taken as the freezing point of
The operation is the water. repeated several times and the average of the observations
taken as the
final value.
The
re-
same operations are now
Fig.
157. Beckmann's apparatus for determining depression of freezing point.
peated after introducing through A' known weights of the sugar be examined (1 to 5 " 100 gms. of water), the maxi.
mum
point to which the mercury rises after overcooling being taken as the freezing point of the solution. The corrected difference between the freezing point of water and that of water sugar is the depression
of freezing point.

was
329
Molecular Depression of Freezing Point. According to what said under osmotic and vapor pressure, solutions of undissociated substances (non-conducting solutions), which contain the same numshould show the same depression of freezing point. gm. mol. of undissociated substance per 1000 gms. of solvent, according to van't Hoff,* is expressed by
ber of gram-molecules per
liter,
The
depression for 1
the formula
002
T2
>
in
which
T is the absolute
of melting
temperature of melting,
This expression in 80 calories and temper2
and
W the latent heat of melting for the solvent.

whose latent heat
abs.,
is
case of water,
ature of melting 273
would give
0.002
X 273 =
80
1.86.
Loomis, as
a matter of fact, in the examination of solutions of some 25 different substances obtained a depression in freezing point for 1 gm. mol. to 1000 gms. of water of almost exactly 1.86 C. The following experiments by
Loomis f give the
results of 6 tests
upon maltose.
342.)
(M, the molecular
weight of maltose anhydride

Grams maltose
grams water
to 1000
(P).
C^HÔu =
330
SUGAR ANALYSIS
A and
Substituting the values obtained for the
of fructose
we
obtain
which agrees closely with the value 180, required by the formula
Hi 2
6.
the weight of water, the If w is the weight of sugar taken and various steps of the calculation are represented by the general equation
:
X 1.86 M= w X 1000 JFXA

The method
is
of determining molecular weight
by the depression
of
one that requires considerable care in manipulation, freezing point and the inexperienced chemist should thoroughly test the method upon substances of known molecular weight before applying it to the examination of unknown compounds. The method is open to a large number
of experimental errors, such as too low a temperature of freezing bath, too high a room temperature, radiation of heat from the observer, faulty thermometer or error in reading, solution of air by the water, For a thorough discussion careless handling of the instrument, etc.
of these various points the chemist is referred to the original papers by Raoult, Beckmann, Loomis and others.* Owing to the small value
of
A any
The
slight error in its
determination becomes greatly magnified in
the final calculation.
method has been successfully employed by Tollens and others in determining the molecular weights of many sugars. The following examples of determinations for nine sugars are selected from a compilation of results by Tollens. f
freezing-point
and Mayer, Brown
arid Morris,
Sugar.

termination.
it
331
Kahlenberg, Davis and Fowler,* for example, have employed measuring the speed of inversion of sucrose. Table LXIII, the above by authorities, gives a comparison of the inversion coefficient
in
by the polariscope and freezing-point methods. One-half gram molecule of sucrose to 1000 c.c. was inverted at 55.5 C. by T&<J gni. mol. of hydrochloric acid.
of sucrose as determined
TABLE LXIII
Giving Rate of Inversion of Sucrose as Determined by Bolariscope and by Depression in Freezing Point
Time.
332
of 0.315
SUGAR ANALYSIS
C. for a solution containing 216.8 gms. of sucrose to 1000 216 8 = 0.634 gm. mols. The elevation in boiling gms. of water, or
'
point for a
is
gm. mol. solution would then be 'OA
0.497 C., which
than the value calculated by van't Hoff's formula. general formula for calculating molecular weights from the elevation in boiling point (A) is similar to the formula for the freezing
slightly lower
The
point
method
(p.
330) and
is
wX
1000
X 0.52
JFXA
The boiling-point method, upon the whole, is open to more sources of error than the freezing-point method and has proved much less satisfactory as a means of establishing the molecular weights of sugars.
CHAPTER
QUALITATIVE
XIII
METHODS FOR THE IDENTIFICATION OF SUGARS

class of organic
PEOBABLY no other
of reactions, or forms so large a
compounds gives such a variety number of chemical derivatives as the
Owing to the great extent of the field it will be possible to sugars. describe only a few of the more general tests and reactions.
for convenience
II.
In describing the various chemical tests, the sugars will be classified under two general groups: I. The reducing sugars.
by when warmed with
The non-reducing sugars. the fact that they cause a
The reducing sugars marked precipitation
are distinguished of cuprous oxide
reducing sugars do not exhibit
Fehling's alkaline copper solution, whereas the nonthis property, or only to a very slight
extent after prolonged boiling. The reducing sugars constitute by far the larger group; of the some one hundred known natural or synthetic sugars, about ninety are reducing and only about ten non-reducing.
Reactions of the Reducing Sugars
The
due
characteristic chemical properties of the reducing sugars are
for the
most part to the occurrence
of a
common
carbonyl-alcohol
H-C-OH
group
I
C=O
I
The reducing
sugars, as aldoses or ketoses, give in
The fact nearly all the reactions peculiar to aldehydes and ketones. to as the chemist must, therefore, first of all, guard against deciding
presence oiâ sugar from a reaction which would also be given by A number of confirmatory formaldehyde, acetaldehyde or acetone. be stated definitely whether it can tests must usually be applied, before
a sugar
or is not present. qualitative reactions for reducing sugars are divided for convenience into I. General tests; II. Special tests; III. Individual
is
The
has been determined from general tests that a sugar is present, special tests must be applied in order to determine what classes or groups of sugars are present, whether hexoses or pentoses, aldoses or
tests.
After
it
After the class or group of ketoses, monosaccharides or disaccharides. must be applied in order tests individual has been sugars ascertained,
333
334
to determine
SUGAR ANALYSIS
what
particular sugars are present.
Only the general
and
special tests are
taken up in the present chapter.
The
individual
II.
tests are given
under the description of the different sugars in Part
GENERAL TESTS FOR REDUCING SUGARS
Among the general tests which are sometimes given for sugars may be mentioned the familiar property which all carbohydrates have of giving off a characteristic sweetish odor upon heating over a flame in a
closed tube.
This odor, which
is
usually designated as caramel-like,
is
however, by many polyatomic alcohols and acids (as by tartaric acid) so that the test is not characteristic of sugars alone. Among the decomposition products obtained by heating sugars in a closed tube
given
off,
may be mentioned (besides water and the gaseous products carbon dioxide and carbon monoxide) formic acid, acetic acid, acetone, furfural and various products of an aldehyde nature. It is to the furfural
and aldehyde products that the characteristic odor of burnt sugar
largely due.
is
The general tests for reducing sugars may be divided for convenience into four general groups of reactions.
I.
II.
III.
Reducing reactions with alkaline solutions of metallic salts. Color reactions with alkalies, acids and phenols. Hydrazone and osazone reactions with phenylhydrazine and
substituted derivatives.
its
IV.
Miscellaneous reactions.
I.
REDUCING REACTIONS OF SUGARS WITH ALKALINE SOLUTIONS OF METALLIC SALTS
sugars and certain of the disaccharides, as maltose and have the property of reducing alkaline solutions of many metallic salts, such as those of copper, silver, mercury and bismuth. 'This reaction, which is common to most aldehydes, is due to the withdrawal of oxygen from the metallic base, the latter being precipitated
lactose,
The simple
either as a suboxide or in the metallic form.
The aldehyde group
of
the sugar molecule is oxidized by the oxygen withdrawn from the metallic base to the acid carboxyl group, as indicated by the following general equation:
H-C:O
Aldehyde
2CuO
Copper Oxide
H-O-C:O
Acid
Cu
O.
Copper Suboxide.
The above, however, marks only the beginning
of the reaction, for, upon heating, the oxidation of the sugar molecule usually proceeds with the

for glycol
335
conversion of alcohol into carboxyl groups as in the following reaction
aldehyde
H-C-O-H
H-C:O
Glycol aldehyde
3Ag
0:C-0-H O:C-O-H
Oxalic
acid
6Ag
Metallic
silver
+ H O.
2
Silver oxide
Water,
This oxidation in the case of the higher monosaccharides is usually attended by a breaking down of the carbon chain as by the oxidation of
glucose in ammoniacal silver solution
:
C
The
12
+ 9 Ag O
2
= 3(COOH)
+ 18 Ag + 3 H 0.
2
reaction between sugars and alkaline salts of metals, as ordinarily carried out, gives rise to a number of monobasic and dibasic acids (formic, oxalic, etc.) in varying proportions according to the conditions
It is not possible, therefore, to express the reaction of the experiment. .by chemical equations except in a very general way. The most common of the alkaline salt solutions employed in test-
ing sugars are those of copper. The sulphate and acetate of copper are the salts most generally used and sugar literature is filled with
will
descriptions of modifications for be described.
making the
test.
Only a few of these
This is the most common chemical Fehling's Copper Solution. reagent employed in testing sugars. As ordinarily prepared the reagent consists of two solutions: solution A containing 34.64 gms. crystallized
c.c. and solution B containing 173 gms. Rochelle and 51.6 gms. sodium hydroxide to 500 c.c. The solutions are the same as those used in quantitative analysis and are to be kept separate until just before using. By mixing 5 c.c. each of solutions A and B in a test tube, adding a few c.c. of the solution to be examined and heat-
copper sulphate to 500

salts
ing to boiling for 2 minutes, a brick-colored precipitate of cuprous oxide, Cu2 0, will form, if reducing sugars are present, the intensity of coloration
and amount
present.
The
of precipitate being proportional to the amount of sugar test is sensitive to about 0.01 mg. of glucose to 1 c.c.
Products Obtained by Heating Reducing Sugars with Fehling's Solutions. The chemical reactions which take place in the oxidation of
Nef,* sugars by means of Fehling's solution are exceedingly complex. who has made the most complete studies in this field, found that in case of 1-arabinose, the oxidation proceeds along three separate lines.
*
Ann., 357, 214-312.
336
I.
SUGAR ANALYSIS
From
10 to 25 per cent of sugar are oxidized to form pentonic
acids.
C Hi
5
= C Hi
5
6.
II.
From 35
to 45 per cent of sugar are oxidized to form formic and oxybutyric acids.
tri-
C H 10
5
+ 2O = HCOOH + C H
4
5.
III.
From 30
to 38 per cent of sugar are oxidized to form formic
and
glycollic acids.
C Hio0
5
+ 30 = HCOOH + 2 C H
2
3.
In case of the hexose sugars, d-glucose, d-mannose and d-fructose, Nef obtained analogous reactions with formation of carbonic, formic, The amount of the glycollic, glyceric, trioxybutyric and hexonic acids. different acids was found to vary according to the amount of alkali
present.
In testing solutions containing
much
urine, the reaction with Fehling's solution
foreign organic matter such as may be interfered with.
Uric acid, creatine, creatinine, albumin, peptones and other substances
may either check the precipitation of cuprous oxide, when reducing sugars
are present, or in
some
plete absence of sugars.
cases cause a precipitate of copper in the comSolutions containing xanthine bases, such as
etc.,
low-grade molasses, distillery waste,

solution
when heated with
Fehling's
greenish-yellow copper compounds, which may be mistaken for cuprous oxide. In all such cases the impure solution
may precipitate
should be clarified with a

excess of lead
is
little normal acetate of lead and filtered any removed from the filtrate with sodium carbonate and
;
the clear solution tested with Fehling's reagent in the usual way. Filtering the- impure solution through animal charcoal is also of ad-
vantage when foreign coloring matter masks the reaction. Barfoed's Copper Solution. Instead of the sulphate, solutions of other copper salts have been employed in testing for sugars. Barfoed * has prepared a solution containing one part crystallized neutral
copper acetate in 15 parts of water; 5 c.c. of 38 per cent acetic acid are c.c. of the copper-acetate solution before use. On boiling the solution a basic acetate of copper is formed, the liberated cupric oxide being reduced in presence of monosaccharides. Barfoed's reagent
added to 200
is
maltose, and
not reduced to any great extent by the disaccharides, lactose and is, therefore, of value in distinguishing these sugars from monosaccharides.
*
Z. analyt.
Chem.,
12, 27.

Soldaini's Copper Solution.
also
337
Carbonate of copper solution has

Soldaini * has prepared a solution
,
been used in testing for sugars.
containing 15 gms. precipitated copper carbonate, CuGO 3 and 416 gms. potassium bicarbonate, KHCO3 dissolved to 1400 c.c. Instead of starting with copper carbonate, copper sulphate may be used; a solution of
,
is added to the KHCOs solution, the precipitate of CuCOs formed being dissolved in the excess of bicarbonate. A solution containing 3.464 gms. copper sulphate and 297 gms. potassium bicarbonate to 1000 c.c. is especially adapted for detecting small amounts of
the latter
first
reducing sugars. Among other copper solutions recommended for testing sugars may be mentioned copper ammonium tartrate and ammoniacal copper sulphate or acetate. None of these preparations has been found, however,
to equal Fehling's reagent for general usefulness in practical
sugar analysis. Tollens's Silver Solution.

tions for detecting sugars
is
The most sensitive of metallic-salt soluammoniacal silver solution, first employed
by Tollens f and hence usually known as Tollens's reagent. This is prepared by dissolving one part silver nitrate in 10 parts of water; a second solution is then made containing one part sodium hydroxide in 10 parts of water. Before making the test equal parts of the two solutions are mixed and then ammonia added drop by drop until the precipitate of
silver oxide is
glucose in 1000 parts of

utes.
completely dissolved. A solution containing one part of water will cause a strong reduction of Tollens's
reagent in the cold, a mirror of silver being deposited within 15 min-
will also
solution containing one part glucose to 100,000 parts of water produce a perceptible reduction. in the cold, but the solution
must stand one to two days. The reduction takes place more rapidly upon warming, but warming or heating the solution is to be avoided owing to the danger of forming explosive silver compounds. For the
latter reason the reagent should
be prepared only just before using. Tests should be carried out in the dark and solutions containing the reagent should not be kept for any length of time.
Tollens's silver reagent is also reduced by all aldehyde substances; affected not only by the sugars which reduce Fehling's solution but also by sucrose, raffinose and all other soluble carbohydrates.
it is
Even the
for
alcohol derivatives of the sugars produce reduction, glycerol, example, causing the formation of a silver mirror. The readiness
is
with which ammoniacal silver solution

*
reduced by soluble organic
Z. Ver. Deut. Zuckerind., 39, 933; 40, 792.
t Ber., 15,
1635; 16, 921.
338
SUGAR ANALYSIS
non-sugars has proved a serious objection against the use of this reagent
in ordinary analytical work.
A third reagent which has been used Knapp's Mercury Solution. Knapp's* alkaline mercuric-cyanide solution. The latter contains 10 gms. of mercuric cyanide dissolved in 100-c.c. sodium
for testing sugars is
hydroxide solution of 1.145 specific gravity. Similar alkaline solutions have been prepared by Sachsse f from mercuric iodide and by Bauer J
from mercuric chloride.
These solutions are reduced upon warming
with sugar solutions giving grayish deposits of metallic mercury. The mercury solutions have the same objection, however, as those of
silver in
creatinine
being reduced by different organic non-sugars, such as creatine, and glycerol and even under certain conditions by alcohol.
Alkaline solutions of mercury salts are, therefore, of but little value in detecting sugar in urine and other liquids rich in organic non-sugars. A fourth reagent, which has been Nylander's Bismuth Solution.
used considerably for detecting reducing sugars in urine,
is
an alkaline
solution of bismuth sub-nitrate, known as Nylander's (or Almen's) This solution as prepared by Nylander is made by dissolving reagent. 2 gms. of bismuth sub-nitrate and 4 gms. of Rochelle salts in 100 gms.
of 8 per cent
the solution
in
sodium hydroxide solution. After standing for a few days filtered through glass wool and the clear filtrate preserved a stoppered bottle. The solution will keep indefinitely. When
is
Nylander's reagent is heated with a solution containing reducing sugars a precipitate of dark metallic bismuth is produced. Heating with TV its
volume
ing.
of 0.01 per cent glucose solution will cause a perceptible darkenIn testing urine 1 c.c. of the reagent and 10 c.c. of urine are heated in a test tube 2 to 5 minutes over the flame; after standing for 5 minutes the solution is examined for the appearance of a dark-
colored sediment.
is open to the same objections noted and mercury solutions. The presence of albumin, nuclein, glucuronic acid and other organic non-sugars in urine will also cause a precipitation of bismuth, even when glucose is comWhile the failure of a precipitate with Nylander's repletely absent.
Nylander's reagent, however,
for the alkaline silver
indicate the absence of reducing sugars, the occurrence of a precipitate may be said to indicate the presence of sugar only when re-
agent
may
ducing non-sugars are proved to be absent.

*
Z. analyt.
Chem.,
9,
395.
Ver. Deut. Zuckerind., 26, 872. t Landw. Vers.-Stat., 36, 304. Z. physiol. Chem., 8, 175.
t Z.

Miscellaneous Solutions.
salts
339
proposed phate and tartaric acid which gives a dark-red precipitate of nickel suboxide in presence of reducing sugars, and alkaline ferric chloride and sodium tartrate which gives a brown-colored precipitate on heating
for sugar testing
Of other alkaline solutions of metallic may be mentioned alkaline nickel sul-
None of these reagents, however, or any of the other alkaline solutions of metallic salts previously mentioned, has been found to equal Fehling's copper reagent for all-around usefulness and
with reducing sugars.
reliability.
II.
COLOR REACTIONS OF SUGARS WITH ALKALIES, ACIDS AND PHENOLS
As a second general reaction of reducing sugars may be mentioned certain color effects which nearly all soluble carbohydrates give when brought into contact with different reagents. The reagents employed
may
be divided into three groups:

I.
Alkalies.
II.
Concentrated mineral acids.

Phenols.
All reducing sugars and alkaline
III.
Color Reactions of Sugars with Alkalies. have the property of coloring solutions of the
alkalies
earths yellow, the application of heat turning the color a dark brown. This reaction is common to all aldehydes. The exact nature of the coloring matter formed by the action of alkalies upon sugars in solution
is
not understood.
Considerable oxygen
is
absorbed from the
air
dur-
ing the reaction and a variety of products of an acid nature are the substances formed.
among
acid
Products Obtained by Heating Reducing Sugars with Alkali. Lactic is produced in considerable amount by the action of alkalies upon
many
reducing sugars such as xylose, arabinose, glucose and fructose.
The presence of calcium lactate in certain sugar-cane molasses is explained by the action of an excess of lime during clarification upon the reducing sugars of the juice. Formic, acetic and oxalic acids have also
been found among the products resulting from the action of alkalies upon sugars in solution. Certain phenol bodies such as pyrocatechin and protocatechuic acid have also been detected among the oxidation products of sugars resulting from treatment with alkalies. Nef * has studied the action of J normal sodium hydroxide upon different sugars
and obtained
in case of d-glucose,
*
d-mannose, and d-fruc-
Ann., 376, 1-119.
340
tose a yield of
SUGAR ANALYSIS
from 40 to 45 per cent
d,l-lactic acid,
from 10 to 15 per
cent d,l-l-hydroxybutyrolactone, about 25 per cent of saccharin, metasaccharin and isosaccharin and a small quantity of tarry decomposition
products. The action of dilute alkalies in causing transformations of sugars into one another by molecular rearrangement is referred to elsewhere.
Color Reactions of Sugars with Mineral Acids.
Treatment
of
solutions of sugars and carbohydrates with concentrated mineral acids gives rise to a number of decomposition products, the color of which
some light upon the nature of the sugars present. most commonly used for this purpose .are sulphuric and The character of the color generated will depend partly hydrochloric. upon the kind of sugar, partly upon the strength of acid used and
frequently throws
The
acids
partly upon the temperature of the reaction. Products Obtained by Heating Sugars with Acids.
The darkening sugar solutions upon warming with concentrated sulphuric or hydrochloric acid is due largely to the formation of insoluble so-called "humus" substances of relatively high carbon content (C = 62
produced in
all
= 3.5 to 4.5 per cent), the percentage of carbon and to 67 per cent and depth of color increasing with the strength of acid used. Attempts have been made to classify the humus substances formed by the action of
acid
upon sugars
into ulmin
and humin and ulmic and humic
acids,
to which various formulae have been assigned by different authorities. The constitution of the humus substances has not been definitely settled, however, and until considerable more work has been done the
formulae of these
must remain more or less a matter of conjecture. In addition to the insoluble humus substances a number of soluble
volatile products are
and
chloric acids
formed by the action of sulphuric and hydroupon sugars. Among such products may be mentioned
formic acid, levulinic acid, furfural, methylfurfural, oxymethylfurfural and a number of dextrin-like condensation or reversion products of high
specific rotation.
The nature and amount of these various products depend largely upon the kind of sugar, and a number of methods of group distinction are based upon the separation of characteristic decompositfon products. Further reference will be made to these under
the special reactions. The ketoses are much
acids than the aldoses
more easily decomposed by strong mineral and their solutions give rise to color reactions with corresponding greater facility. This offers one means of distinguishing between a ketose and aldose or of detecting a ketose sugar in presence of an aldose. If a cold sugar solution be treated in a test
341
tube with a few cubic centimeters of concentrated sulphuric acid, allowing the latter to flow down the walls of the tube to the bottom without
shaking, a
brown
if
sugar solution
ring will quickly form at the junction of the acid and fructose, sucrose or a sugar containing the ketone
with glucose, lactose, maltose and the aldoses in is present; general no such coloration will develop. Color Reactions of Sugars with Phenols. The most distinctive
group
color reactions of the sugars are those obtained
by treatment with
different phenols in presence of concentrated hydrochloric or sulphuric acid. The development of a color in this case is due to the formation
of condensation products between the phenol derivatives and the decomposition products obtained from the sugar (humus substances,
furfural, aldehydes, etc.). a-Naphthol, thymol, resorcin, orcin, naphthoresorcin and phloroglucin are among the more important phenol derivatives used for making color reaction with sugars.
phenols are performed in various with for ways. a-naphthol, example, which is perhaps used more frequently than any of the others, is made as follows: 1 to 2 cubic centimeters of the sugar solution are treated in a test tube with 1 to 2 drops of a 10 to 20 per cent alcoholic solution of a-naphthol. A few cubic centimeters of concentrated sulphuric acid (must be free from
The
color reactions with the

test
The
nitric acid) are
then carefully added so as to flow down the walls of the tube to the bottom. If sugars containing a ketone group are present a violet ring will form instantly at the junction of the two liquids; in
presence of aldoses a gentle warming of the test tube sary in order to bring out the full intensity of color.
is
usually neces-
The a-naphthol
test, which is of extreme delicacy, is frequently employed in sugar houses and refineries in testing the condensation water from the vacuum pan for presence of sucrose lost by entrainment. If the reaction described for a-naphthol is carried out with thymol,
menthol, resorcin and other phenols similar colorations are produced, the tints varying from cherry red to deep purple. The tests with phenols and hydrochloric acid are usually made by warming a few cubic centimeters of the sugar solution with a solution
of the phenol (resorcin, orcin, phloroglucin, etc.) in concentrated hydrochloric acid. The colorations thus obtained are usually very brilliant,
varying in tint from a bright red to a bluish violet. The colors formed are not permanent, however; they rapidly darken and the clear-colored solution soon becomes turbid with the precipitation of a dark-colored
condensation product.
342
SUGAR ANALYSIS
USE OF THE SPECTROSCOPE IN STUDYING COLOR REACTIONS FOR SUGARS
The spectroscope has been used with great

his
success by Tollens and coworkers in studying the colors obtained by treating sugars with
different reagents.
in different parts of the spectrum,
The appearance of characteristic absorption bands when the colored solution is viewed
light, is peculiar of many sugars. Direct-vision A simple type of Spectroscope. Description for the is direct vision inabsorption studying spectra spectroscope
through the spectroscope against white

of
Fig. 158.
3-
Showing outer and inner construction
Fig. 159. of a direct-vision spectroscope.
strument illustrated in Fig. 158, the interior construction of which
is
shown in Fig. 159. The essential parts
of the apparatus consist of a telescopic tube conand an achromatic objective 0. At one end taining an Amici prism of the tube, protected by the screw cap K, a diaphragm is situated con-
343
taining a narrow slit S, the width of which can be adjusted by turning the milled ring B. The upper half of the slit is covered with a small
prism V; a mirror D, which can be rotated through a small angle about the axis of the tube, is also attached to the slit end of the instrument.
tube
of the spectroscope there is fixed a small lateral a graduated scale E. The latter is attached to a containing small prism b to which is fixed a converging lens a. At R is a right
At the prism end
angle prism, from the hypotenuse surface of which the image of the is reflected through the achromatic objective 0' upon the cut scale
surface cc of the
If
the
slit
Amici prism. end of the spectroscope be pointed towards a sodium

1,
2 and
flame the rays of light will pass into the spectroscope along the paths 3. The telescope is first focused by turning the milled ring
until a sharply defined
is
obtained by the light
image of the lower uncovered half of the slit passing along 2 upon the surface cc. The image
of the scale
is
reflected at the
same
time,
by the
light passing along
3 ; also
upon
cc.
The
position of the
sodium
line is
noted upon the
If the the latter being in this way standardized. spectroscope be now directed towards the sky a continuous spectrum is next turned until the is obtained upon the surface cc; the mirror
graduated
scale,
upon through an opening in the cap the small prism V and thence through the upper half of the slit S', in this way a continuous spectrum is obtained upon cc the width of which
light passing
along
1 is reflected
equal to the total length of the slit S. If the slit has been sufficiently reduced in width, the spectrum of sunlight is seen to be crossed by a number of dark lines, the so-called
is
Fraunhofer
light
of
lines, which are due to the absorption of certain rays of from the incandescent mass of the sun by the vaporized elements the solar atmosphere. A dark line (the D line of Fraunhofer's scale),
for example, corresponds to the position of the bright-yellow line obtained with the sodium flame and so of the other elements. The
position and wave-length of the more important Fraunhofer lines is shown in Fig. 165 (p. 384) their presence is very helpful in defining the
;
position of absorption spectra.
For studying absorption spectra the spectroscope is mounted upon a stand as shown in Fig. 160, a screen L being attached to the tube to shade the eye of the observer. The solution to be examined is placed in a small cell T, before the front opening in the screw cap and viewed The rays of light absorbed by the solution will against white light. cause characteristic dark-colored bands to appear upon that part of the
spectrum corresponding to the lower half of the
slit.
The part
of the
344
SUGAR ANALYSIS
covered by the prism V spectrum corresponding to the half of the slit meanwhile remains continuous and together with the scale, or Fraunhofer lines, serves for the exact location of the absorption bands.
_...
::
Fig. 161. Fig. 160. Methods of mounting apparatus for study of absorption spectra.
Solutions which arc only weakly absorptive are best examined through a large tube H, in the manner shown in Fig. 161. The spectroscope is turned and clamped in a vertical position and the light reflected upward
from the mirror

Tollens's
through the glass bottom of the support G. In preparing of Studying Absorption Spectra. color tests of sugar solutions for spectroscopic examination it is important that the color remain permanently in solution and that no turbidity develop which would obscure the visible parts of the spectrum. This is sometimes accomplished by carrying out the reaction in presence of alcohol or some other solvent to hold the color compound in solu-
Method
tion.
better
methode ")
way is by use of Tollens's* deposit method ("AbsatzIn this method the deposit of insoluble condensation prod*
Ber., 29, 1202.

ucts obtained
345
by treating the sugar solution with hydrochloric acid and the phenol (orcin, phloroglucin, naphthoresorcin, etc.) is filtered off, washed several times with water and then dissolved in alcohol. Brightcolored solutions are thus obtained which can be brought by dilution with
alcohol to the degree of intensity suitable for spectroscopic examination. Descriptions of characteristic absorption spectra will be given under the
reactions for groups
and individual
sugars.
importance than the color reactions with phenols are the by treating sugars with aromatic amines (aniline, xylidine, diphenylamine, etc.) in presence of concentrated hydrochloric acid. The colors in this instance are due to a combination between the aromatic amine and the furfural, methylfurfural, and oxymethyl.
Of
less
color tests obtained
furfural derived
from the decomposition of the sugar.
III.
HYDRAZONE AND OSAZONE REACTIONS OF REDUCING SUGARS WITH PHENYLHYDRAZINE AND ITS SUBSTITUTED DERIVATIVES
In many respects the most important of the qualitative tests for sugars are those obtained with phenylhydrazine and its substituted derivatives. Phenylhydrazine was introduced as a reagent in sugar
chemistry by Emil Fischer* in 1884; it has been of immense service not only as a means of separation and identification but also in first
opening a
way
to a thorough understanding of the molecular constitu-
tion of sugars.
Hydrazone Reaction.
The
reaction
with phenylhydrazine
is
limited to such sugars as contain a free carbonyl group and proceeds in two phases with production of two entirely different classes of com-
pounds.
The
first
and ketones, the of the carbonyl group combining with H 2 of the amino group in the phenylhydrazine with formation of a group of compounds called hydrazones. With formaldehyde, for example, the
reaction proceeds as follows:
phase of the reaction
is
common
to
all
aldehydes
H C:0
2
H N-NHC H
2 6
H C:N-NHC H
2
6
HO
2
Formaldehyde
Phenylhydrazine
Formaldehydephenylhydrazone
Water
With the carbonyl group

diose
:
of a sugar the reaction
would be
for a
CH OH
2
CH OH
2
HC:0
Diose
H N-NHC H
2 6
HC:N-NHC H
6
HO
2
Phenylhydrazine *
Dioae-phenylhydrazone
Water
Ber., 17, 579.
346
SUGAR ANALYSIS
reaction is carried out by treating the sugar solution with a solution containing one volume of phenylhydrazine, one volume of 50 per cent acetic acid, and three volumes of water. A little more of the phenylhydrazine is used in making the test than the
in the cold
The hydrazone
present.
theoretical quantity corresponding to the supposed amount of sugar In place of the above solution the crystalline chloride of
phenylhydrazine may be used to advantage, a few grams of sodium After the above acetate being also added to promote the reaction. treatment the hydrazones of the sugars will separate sooner or later as well-defined crystalline compounds, the length of time for separation depending upon the solubility of the hydrazones formed. The phenyl-
hydrazone of mannose, for example, being very insoluble, will separate almost immediately; those of the methylpentoses, fucose, rhamnose and rhodeose also deposit readily; the phenylhydrazone of glucose, on the other hand, which is quite soluble in water, may require one or two
days for its precipitation. By filtering off the hydrazones as they are formed a separation of sugars in mixtures may often be accomplished. After separation of the hydrazones the latter are filtered off and recrystallized either from water or, in case of difficultly soluble hydrazones, from alcohol or pyridine. In place of Use of Substituted Derivatives of Phenylhydrazine. phenylhydrazine any of its substituted derivatives may be used for the
purpose of precipitating sugars.
yield in
many
substituted phenylhydrazines cases characteristic hydrazones with sugars and their use
The
in sugar chemistry in recent years has
been of the greatest service. Of the various substituted phenylhydrazines the following are among the
most important.
1.
Methylphenylhydrazine
HN NX
2
2.
Ethylphenylhydrazine
HN Nx N
2
CH
6
3.
Amylphenylhydrazine
HN NX
2
4.
Allylphenylhydrazine
HN NN
2
C 6H 5 CH
C 6H C
C*
6
5.
Diphenylhydrazine
HN NX
2
H
TT
6.
Benzylphenylhydrazine
/ H N Nx
2
CeH 6

TT
7.
347
Parabromophenylhydrazine
HN N
2
C 6 H 4Br
TT
8.
Paranitrophenylhydrazine
HN N
2
H N0
4
Other hydrazines than those of the phenyl group are also employed
as, for example,
TT
9.
Naphthylhydrazine
H N-N^
2
CloHj
The reactions with the substituted hydrazines are usually best carried out in alcoholic solution, the hydrazones formed being for the most part much less soluble than those of ordinary phenylhydrazine.
In the examination of the hydrazones obtained from sugar solutions a melting point of the product is taken before and after recrystallizaIf the melting point remains unchanged the hydrazone is pure. tion. Should a difference in the temperature of melting be obtained the
hydrazone should be recrystallized until successive determinations show no change in melting point. A table of melting points will then usually identify the hydrazone of the sugar. (See Table 24, Appendix.) When a sufficient quantity Separation of Sugars from Hydrazones. of hydrazone is available it is always well to decompose the compound and make a direct examination of the separated sugar. For the separation of sugars from their hydrazones two processes are available: First, by means of concentrated hydrochloric acid as originally used by
Fischer.
Second, by means of benzaldehyde and formaldehyde as rec-
ommended by Herzfeld* and by
Ruff.f the hydrazone of a sugar is treated with concentrated hydrochloric acid the chloride of the hydrazine and free sugar are formed:
When
12
5
N-NHC H5 +
6
I
HC1
+ H
is
H O
12
HC1
H N-NHC H
2 6
Hexose-pheny hydrazone
Hexose
Phenylhydrazine
chloride
The phenylhydrazine
almost insoluble in concentrated hydrochloric acid and is removed by filtration. The filtrate is neutralized with lead carbonate; the lead chloride is filtered off and the filtrate evaporated to a syrup. The latter is shaken with 95 per cent alcohol, any
chloride
remaining lead chloride filtered off and the alcoholic filtrate evaporated to a sirup which is set aside for the sugar to crystallize.
*
Ber., 28, 442.
t Ber., 32, 3234.
348
SUGAR ANALYSIS
hydes
The separation of sugars from their hydrazones by means of aldeis much simpler than by use of hydrochloric acid and this is the
For this purpose benzaldehyde process most generally used at present. is usually employed for the hydrazones of phenylhydrazine and formaldehyde for the hydrazones of the substituted hydrazines. The reaction
between the aldehyde and hydrazone is a simple one, the aldehyde placing the sugar with formation of aldehyde hydrazone.
dis-
+
Hexose-phenylhydrazone
CeHsCHO
Benzaldehyde
12
+ C H CHN-NHC H5
6
6
Hexose
Benzaldehydephenylhydrazone
C Hi O N-N(C H
6
2 5
5) 2
CH O
2
Hi 2 O 6
CH N-N(C H
2
6
5)2
Hexose-diphenylhydrazone
Formaldehyde
Hexose
Formaldehydediphenylhydrazone
The reaction is best carried out by treating a solution of the hydrazone in 50 per cent alcohol in a flask with an amount of the aldehyde slightly in excess of the theoretical quantity necessary to effect de-
The flask is then attached to a reflux condenser and composition. the solution gently boiled for an hour. After cooling, the solution is filtered from the aldehyde hydrazone, the filtrate shaken out several
times with ether in a separatory funnel, the sugar solution, after decolorizing with animal charcoal, evaporated to a sirup and set aside for
Should crystallization not take place immediately, crystallization. the process may be promoted by priming the sirup with a minute After crystallization the crystal of the sugar suspected to be present.
sugar crystals are filtered off, washed with alcohol and ether (using suction) and dried between filter paper in a desiccator over concentrated sulphuric acid. The identity of the sugar thus obtained established by determination of its specific rotation.
is
then
If the filtrate obtained from filtration of a hydrazone be shaken out with ether to remove excess of hydrazine, the solution can be treated a second time with a different hydrazine. In this manner a qualitative
separation of several mixed sugars may be accomplished. Osazone Reaction. While the hydrazone reaction
is
of pre-
eminent value in the isolation of sugars, the osazone test with phenylhydrazine is usually of more qualitative significance owing to the greater insolubility of the osazones in water and the consequent greater rapidity and ease of their separation as compared with hydrazones.
If a solution of a reducing sugar be treated with an excess of phenylhydrazine and then warmed, two molecules of phenylhydrazine unite with the sugar molecule forming an osazone. The aldehyde or ketorffe
349
group of the sugar and the adjacent alcohol group are the ones which always participate in this reaction.
CH OH
2
CH OH
2
(HCOH)s_
(HCOH)
HCOH HC:O
Hexose
+ ^N-NHCeHs
Phenylhydrazine
C:N-NHC H HC:N-NHC H
6 6
+ 2H O
2
H,
Hexose-phenylosazone
Water
Hydrogen
The
of
free
hydrogen liberated
in the
above reaction acts upon a part
of the excess of phenylhydrazine reducing this to aniline with liberation
ammonia.
2 6 5
H N-NHC H
Phenylhydrazine
NH C H
2
NH
Hydrogen
Aniline
Ammonia
Since the first stage in the reaction with phenylhydrazine is the formation of a hydrazone, it follows that all phenylhydrazones when treated with phenylhydrazine in excess are changed to the corresponding osazones.
CH
6
12
N-NHC H + H N-NHC H
6 5
2 6
= C 6 H 10O4(N-NHC
5) 2
+ HO + H
2
Hexose-phenylhydrazone
Phenylhydrazine
Hexose-phenylosazone
Water Hydrogen
In conducting the reaction for osazones the original method of Fischer* is usually followed. For 1 gm. of sugar, 2 gms. of phenylhydrazine chloride and 3 gms. crystallized sodium acetate (CH 3 COONa 3 2 O) and 20 c.c. of water are heated together for f to 1J hours in a large test tube of about 50 c.c. capacity placed in a boiling-water bath.
The contents
tion.
of the tube are stirred occasionally to
promote
crystalliza-
a solution of phenylhydrazine acetate, prepared by adding concentrated acetic acid drop by drop to phenylhydrazine until the turbid emulsion clears. The osa-
Instead of the chloride one
may employ
zone reaction with the substituted hydrazines way as with phenylhydrazine.
is
conducted in the same
The osazones
pounds
crystallize
of the sugars are yellowish-colored crystalline
com-
of variable solubility.
The osazones
of the monosaccharides
and
lactose,
out from the hot solutions; those of the disaccharides, maltose however, separate only after cooling. A separation of the
osazones of the mono- and disaccharides can be accomplished in this manner, a second crystallization usually rendering the separation com-
While the osazones of the monosaccharides are nearly all of much lower solubility than the corresponding hydrazones, the osazone
plete.
separation
is
never complete.
*
Ber., 17, 579.
350
Yield and
SUGAR ANALYSIS
the
Time for Formation of Osazones. Sugars differ greatly in amount of osazone which is formed under a definite method of treatment, and this property has been utilized as a means of identifica-
Maquenne,* for example, has determined the yield of osazones obtained by heating 1 gm. of different sugars in 100 c.c. of water with 5 c.c. of a solution, containing 40 gms. phenylhydrazine and 40 gms. The glacial acetic acid in 100 c.c., for 1 hour in a boiling- water bath.
tion.
sugars studied by Maquenne are arranged in Table of yield of osazone.
LXV
in the order
TABLE
Showing Yield of Osazones and
LXV
of Precipitation for Different Sugars
Time
Sugar.
351
small test tube, corked loosely to prevent evaporation and heated in The tube is shaken occasionally without removing from boiling water. the bath and the time noted for the separation of a precipitate. Under the above conditions Mulliken noted the following:
Sugar.
352
SUGAR ANALYSIS
Influence of Lactose on Glucose
Weight
of
glucose.

Influence of Sucrose on Fructose
353
Weight
of fructose.
354
alkalies) give the
SUGAR ANALYSIS
same osazone.
This
is
made more
clear
from the
fol-
lowing stereoformulse of glucose,
mannose and
fructose.
H-C=0 H-C-OH
I
H-C = HO-C-H
I
CH OH C=O
2
HO-C-H'"
"HO-C-H"
H-C-OH H-C-OH
CH OH
2
H-C-OH H-C-OH
CH OH
2
HO-C-H H-C-OH H-C-OH

CH OH
2
d-Glucose
d-Mannose
d-Fructose
The part
dotted line
of the molecule
all
below the dotted
line
has the same spatial
arrangement in
is
three sugars. The part of the molecule above the the only part of the molecule affected in the osazone re-
action, this in all three sugars giving rise to
an osazone which has the
same
structural formula:
H-C=N-NH-C H
6
HO-C-H H-C-OH H-C-OH

This circumstance, although nullifying the use of phenylosazones in certain cases as a means of identification, has yet thrown a flood of
light
upon the molecular constitution of sugars. Test for Ketoses with Methylphenylhydrazine.
In distinction from
phenylhydrazine the substituted hydrazines do not always give the same osazone reaction with sugars which are mutually transformed. The osazone reaction with substituted hydrazines has, therefore, a dis-
Methylphenylhydrazine, for example, forms very readily a characteristic osazone with d-fructose, but does not form an osazone with d-glucose or d-mannose or any of the other aldose
tinct qualitative value.
sugars.
The osazone
fore, serviceable in distinguishing aldoses
reaction with methylphenylhydrazine from ketoses.
is,
there-
While hydrazones, upon Decomposition of Osazones into Osones. decomposition with strong hydrochloric acid or with benzaldehyde or formaldehyde, yield the component sugar, the osazones cannot be resolved in this manner. The osazone reaction is consequently of value
355
only as a means of identifying and not of separating sugars. The decomposition of osazones with acids and aldehydes has, however, a considerable theoretical interest which may be considered briefly in this
connection.
Treatment
of osazones with concentrated hydrochloric acid or with
certain aldehydes causes, as in the case of hydrazones, a separation of
the phenylhydrazine the product remaining behind, however, is not the original sugar, but a compound with two adjacent carbonyl groups called
;
an osone.
ample,
2
The
reaction of glucosazone with hydrochloric acid, for ex-
is:
CH OH
(CHOH) 3
CH OH
2
+ 2 HC1 + 2 H O
2
(CHOH)
+ 2 CeHsNH - NH HC1
2
C N - NHC H HC N - NHC H
:
C O
:
HC O
:
Glucosazone
Hydrochloric acid.
Glucosone
Phenylhydrazine chloride.
In case of osazones soluble in hot water the conversion into osones can be easily effected with benzaldehyde in presence of sufficient alcohol, the phenylhydrazine being separated as benzaldehyde-phenylhydrazone and the osone remaining behind in solution.
Osones upon treatment with zinc dust and acetic acid are reduced by the nascent hydrogen to a sugar, the end carbonyl group being converted always to an alcohol group, as shown in the following equation
for glucosone.
CH OH
2
CH OH
2 3
(CHOH)
(CHOH) 3
C:O
C:0
HC O
:
CH OH
2
Glucosone
Hydrogen
Fructose
be seen from the above reaction that the sugar obtained by reduction of an osone is always a ketose. By this means glucose and mannose can be transformed into fructose, and this type of reaction is
It will
true for the conversion of
any aldose
into the corresponding ketose, the
steps of the transformation being always
Aldose
Osazo'ne
>
Osone
>
Ketose.
The
different sugars to
osones, while of great service in establishing the relationship of one another, have no value either in qualitative or
quantitative sugar analysis.
356
SUGAR ANALYSIS
THE IDENTIFICATION OF HYDRAZONES AND OSAZONES
The
identification of
hydrazones and osazones, by examination of
their physical properties, although belonging strictly to the tests for
individual sugars,
is
introduced for convenience at this point.

of melting point identification of
is
Determination of Melting Point of Hydrazones and Osazones.
The determination
for
the principal physical method
hydrazones
and
capil-
osazones.
Capillary-tube
Method.
is
The
lary-tube
ally
method
the one most gener-
employed
for
determining melting
in
points. way of apparatus are
The
essential requirements
shown
is
in Fig. 162.
long-neck flask with a
small
filled
body
about
of about 20-c.c. capacity
two-thirds with pure concentrated sulphuric acid; to prevent discoloration of
the acid through accidental contamination with organic matter a small crystal
Fig.
of potassium nitrate, the size of a pinAp- head, is dropped in. The flask is clamped paratus for de- ^ o a lamp-stand in the manner shown.
162.
The opening
of the flask
is fitted
with a
perforated cork containing a groove upon the side to allow escape of expanding air. The perforation in the cork should be of such a size as to hold a
thermometer, graduated to 300 C., tightly in position; the bulb of the latter should be above the bottom of the flask and yet be submerged entirely in the acid.
The capillary tubes for holding the hydrazone or osazone are best prepared by thoroughly softening a piece of glass tubing by turning it in the flame and then drawing it out to about 1 to 1.5-mm. diameter. By
number
continuing this process backwards along the tube a Fi g- 163 ing of sections are obtained similar to Fig. 163a; the sections are then filed off at the points indicated
-
Show-
and the smaller ends melted together in the flame, melting points. Small tubes of the size and shape shown in Fig. 1636 are thus obtained. A small amount of finely powdered hydrazone or osazone is then introduced into the open end of the tube and the latter gently tapped until

the substance has settled to the bottom.
357
To
prevent the powdered
material from forming too loose a layer it is usually well to push it The depth tightly down by means of a platinum-wire or thin-glass rod. 2 mm. not exceed The in the tube should of substance capillary tube
containing the substance is then attached to the thermometer either by binding it with a piece of fine platinum wire or by dipping it first in concentrated sulphuric acid and allowing it to stick to the thermometer
bulb by adhesion. The tube is placed so that the layer of substance even with the center of the mercury bulb.
is
After placing the thermometer and tube in position, as shown in Fig. 162, a small flame is placed beneath the flask and the temperature raised
until the liquefaction of the powdered crystals indicates the temperature of melting. Hydrazones and osazones at the point of melting dewith compose darkening of color, the evolution of gas causing the
liquefied substance to
foam upwards
in the
stem of the tube.
The
first
determination of melting point is only preliminary and a second and The third trial should always be made with fresh tubes and material.
acid in the subsequent tests is heated rapidly to about 5 C. below the melting point first observed and then the temperature raised
gradually so that the thread of mercury in the thermometer comes to rest just at the point of liquefaction. The entire operation for glucosa^
zone, for example, melting at 204 to 205 C., should not consume over 4 minutes. Undue protraction of the time of heating affects the result of the determination very markedly and the wide discrepancies noted in the literature between melting-point determinations of the same osazone by different authorities are due largely to this cause. A second method for determining melting Maquenne's Block. of is employed considerably by French and osazones hydrazones points chemists. This method involves the use of the Maquenne Block, an apparatus invented by Maquenne in 1887, the essential features of which are shown in Fig. 164.
The important part of Maquenne's apparatus consists of a prismatic block (A) of brass, weighing about 2 kilos, which is placed in a frame with one of its edges resting above the openings of a long gas burner (B).
In one end of the block about 5
mm.
below the upper surface a hole
is
bored, extending nearly the length of the block, into which a thermometer (T) can be inserted. In the upper level surface of the block are a
number of small, round cavities. In conducting a determination a small amount of substance is placed in one of the cavities, which, to prevent disturbances from air drafts, is covered with a small glass; the thermometer is then inserted so that its bulb is about underneath the cavity and
358
SUGAR ANALYSIS
the burner started with a low, uniform flame. The temperature is slowly elevated until the substance begins to melt when the thermometer
drawn out or pushed in until just the end of the mercury thread proand the temperature noted. The block is now cooled slightly and a second determination made more slowly than before, using a cavity above the bulb of the thermometer in its second position. Owing to the fact that the block has nearly the same temperature, the entire column of mercury is brought to the same temperature as that of the melting substance and no correction due to contraction of the thread outside the unheated portion of the thermometer is necessary as by the method of melting-point determination previously described.
is
jects
Fig. 164.
Maquenne's block
for determining melting points.
comparison of melting points of glucose-phenylosazone by the
two methods shows the following: capillary tube 205 C. (Fischer), Maquenne Block 230 to 232 C. (Bertrand). From this it would appear that the Maquenne Block gives considerably higher melting points than the capillary-tube method. A critical comparison of the two methods by Miither * (see Table LXVI, opposite page) shows, however, that this is not
It will
always the case. be seen that Miither obtained for glucosazone results by the block agreeing very closely with those by the tube, the range found by the block being 200 to 206 C. and by the tube 203.5 to 205 C. The greater variation by the block is attributed by Miither to the
unequal distribution of heat through the brass, the outer surface being more quickly warmed than the center; differences from 3 to 6 C. were also noted for different positions of the thermometer inside the block. The slowness with which the block is heated and cooled and
the difficulty with which the cavities are cleaned are also serious objections. With substances which sublime, the Maquenne Block cannot
*
Dissertation, Gottingen, 1903.
359
be used on account of the rapid condensation of material from the cavity upon the cover glass. These objections together with the high
cost of the apparatus (about $15.00, duty free) render it much less desirable for determining melting points than the simpler capillary-tube
method.
TABLE LXVI
Showing Melting Points of Hydrazones and Osazones by Different Methods. (Miither.)
Compound.
360
SUGAR ANALYSIS
attributed to the existence of hydrazones of abut the conditions for their separate formation have not
The isomerism has been

and
jS-glucose,
been definitely established. Similar differences have been noted in the case of other hydrazones, but whether the variation in properties is due to isomerism or to a difference in purity is not always certain.
Optical Activity of Hydrazones and Osazones as a Means of IdenIn addition to melting point the optical activity of hydrazones and osazones is sometimes employed as a means of identification.
tification.
solutions the polarization of hydrazones and osazones can not always be measured with exactness. In the case of
color of
Owing to the low some of the
solubility of
some
of the
compounds and the high
hydrazones the existence of different isomers, as in the case of glucosephenylhydrazone just cited, may cause wide differences in polarization.
Mutarotation, which was noted in the case of glucose-phenylhydrazone, has also been observed with some of the osazones. Thus Allen and
Tollens* found for 1-arabinose-phenylosazone [a]o =+18.9 after dissolving in alcohol, but after standing a short time the solution became
optically inactive.
The rotatory power

for different solvents.
of hydrazones
and osazones also varies greatly Thus Lobry de Bruyn and van Ekenstein f
found the following rotations for different 0-naphthylhydrazones in methyl alcohol and glacial acetic acid.

TABLE LXVII
Giving Polarization of Different Osazones
361
1-Arabinose-phenylosazone 1-Arabinose-p-bromophenylosazone Xylose-phenylosazone
+028' -015'
Xylose-p-bromophenylosazone Rhamnose-phenylosazone d-Glucose-phenylosazone d-Glucose-p-bromophenylosazone. d-Galactose-phenylosazone Sorbose-phenylosazone Maltose-phenylosazone Lactose-phenylosazone
+048' -015' +130'
The method
rotations are small

of identification
and
in
some
cases uncertain so that this

satisfactory than a melt-
upon the whole
is less
ing-point determination. In case of the hydrazones and osazones of optically opposite isomeric sugars (which, as regards melting point and solubility, behave alike ex-
cept in the special case where optically active hydrazines are used), a determination of the optical activity of the compound is the only ready means of identification. Thus Fischer * gives for the phenylhydrazones of d- and 1-galactose the following constants.
Melting point.
[a]^
d-Galactose-phenylhydrazone
1-Galactose-phenylhydrazone
158
21.6
158
+21.6
Fischer also gives for the phenylhydrazones of d- and 1-mannose
M
'Sf
Station.
d-Mannose-phenylhydrazone 1-Mannose-phenylhydrazone
195
- 1.2
+1.2
195
rotations in the latter case were the angular readings obtained 100-mm. tube upon a solution of 0.1 gm. hydrazone in 1 c.c. cold concentrated hydrochloric acid and diluted with 5 c.c. of water.
in a
The
Employment
of
Optically
Active
Hydrazines
for
Separating
Sugars from Racemic Mixtures.
Neuberg f has recently employed
optically active hydrazines for analyzing racemic mixtures of sugars.

If two optically opposite isomeric sugars (" antipodes ") S and S form hydrazones with an optically inactive hydrazine H, the resulting compounds, which may be represented by the symbols +SH and
SH
are also antipodes, and, although of exactly opposite rotations,

*
Fischer's
"
Untersuchungen uber Kohlenhydrate."

38, 866, 868.
t Ber., 36, 1192;
362
have
SUGAR ANALYSIS
in other respects, such as specific gravity, melting point, solubility, same physical properties. A separation of two such hydrathe etc., zones is consequently not possible by the ordinary methods of analysis.
If,
however, the two sugars
+S
and
S combine with an optically
and S hydrazine as +H, the resulting hydrazones are not optical antipodes and show well-defined differences S A separation of the in solubility, melting point and other properties. two hydrazones is thus made possible by the ordinary methods of
active
+ +H
+H
fractional crystallization.
The
hydrazines, which have been used
by Neuberg and
his co-
workers for this method of separating sugars, are 1-menthylhydrazine and d-amylphenylhydrazine, the structural formulae of which are as
follows
:
CH
CH
in
)cH-CH
2
CH CH-NH-NH
2
\ /
CH
CH -CH-CH
3
l-Menthylhydrazine.
d-Amylphenylhydrazine
The method has been employed
successfully
by Neuberg
in resolv-
ing the racemic sugar d,l-arabinose, which occurs in the urine of
many
persons suffering from pentosuria; d,l-arabinose gives with 1-menthylhydrazine an easily soluble 1-arabinose-l-menthylhydrazone and a very
insoluble d-arabinose-1-menthylhydrazone.
The
is
latter is filtered off
upon treatment with formaldehyde
(p.
348)
easily
and decomposed with
liberation of the free sugar d-arabinose.
IV.
MISCELLANEOUS REACTIONS OF SUGARS
Reactions of Sugars with Reducing Agents. The simple reducing in their of are character or sugars, aldehydes ketones, easily transformed by reducing agents into the corresponding alcohols. The sugar mannose, for example, is reduced by sodium amalgam to the alcohol mannite.
CH OH
2
CH OH
2
(CHOH) 4
(CHOH) 4
CHO
Mannose
CH OH
2
Hydrogen
Mannite
more general type CnH 2n O n

Sugar
of equation
would be:
C n H 2n+2 O n
Sugar alcohol
Hydrogen

The
363
reactions of the different sugars with reducing agents are of comparatively minor importance as regards use in sugar analysis. description of the different sugar alcohols, with reactions and
methods
of identification, is given in Chapter XXIII. Reactions of Sugars with Weak Oxidizing Agents. sugars belonging to the aldoses are changed by means of the
ful oxidizing agents,
Reducing
less
monobasic
2
I
acids.
powersuch as bromine water, into the corresponding Thus:
CH OH
(CHOH)<
Aldo-hexose
CH OH
2
4-
2Br
(CHOH) 4
Hexonic acid
2HBr
Hydrobromic acid
Bromine water
1
In carrying out the reaction

of water
part sugar
is
treated with 5 parts

at
and 2 parts
of bromine,
and the solution kept
room tempera-
ture for 1 to 3 days.
Ketose sugars, upon treatment with bromine water, undergo but oxidation during the first few days. Prolonged action, or elevation of temperature, will, however, oxidize ketoses with a breaking up of the molecule into several acids of fewer carbon atoms. Rate of Oxidation with Bromine as a Test for Aldoses and Ketoses. The rate of oxidation of several aldose sugars with bromine water, as compared with fructose, is shown in the following experiments by Votocek and Nemecek;* 0.5 gm. of pure sugar was dissolved in a 50-c.c. flask in 9 c.c. of water, 40 c.c. of bromine water (saturated at room temperature) were then added and the volume made up to 50 c.c. After standing at room temperature (21 C.) for 24 hours, the unoxidized sugar was determined in each flask with the following results:
little
Sugar.
364
SUGAR ANALYSIS
agents, as 30 per cent nitric acid, into
warming with stronger oxidizing

the corresponding dibasic acids.
Thus
CH OH
2
COOH
(CHOH) 4
2HN0
(CHOH) 4
+ 2H 0+2NO
2
CHO
Galactose
Nitric acid
COOH
Mucic acid
is
In carrying out the reaction one part of sugar

nitric acid of 1.2 sp. gr.
heated with 2| parts

until
and gently warmed at 40 to 50 C.
no
more
nitrous fumes are evolved.

all nitric
The
is
water bath until

the acid or
its
acid
then heated upon the expelled and then evaporated, when

solution
is
lactone will in
many
cases crystallize;
when
crystalliza-
tion does not occur separation from impurities is effected by forming an insoluble salt or other derivative from which the acid can afterward be liberated in the pure condition.
Ketose sugars, upon oxidation with

largest
nitric acid, are

is
degraded into
lower oxidation products, of which oxalic acid
usually formed in
amount.
substituted aldose sugars, as the methyltetroses, methylpenthe methyl group upon oxidation with
The
toses, methylhexoses, etc., lose

nitric acid
and are degraded into dibasic acids

3
of one less carbon atom.
CH
CHOH
(CHOH) 2
COOH
+ 5O
(CHOH) 2
HCOOH
Formic acid
HO
2
CHO
Methyltetrose
COOH
Tartaric acid
Water
way the methylpentoses, rhamnose, rhodeose and fucose are oxidized into trioxyglutaric acids, the methylhexoses into
tetraoxyadipic acids, etc.
In the same
Oxime Reaction of Sugars. Many of the reducing sugars react with hydroxylamine, after the manner of all aldehydes and ketones, with formation of oximes. The following combination of glucose with
hydroxylamine
is
an
illustration of this
type of reactions.
CH OH
2
CH OH
2
(CHOH) 4
(CHOH) 4
+'
H-C:O
Glucose
H N-OH
2
H-C:N-OH
Glucose-oxime
Hydroxylamine
Water
The oximes
of the sugars are often difficult to isolate
and the
reac-
tion, for this reason,
value in sugar analysis. In sugar synthesis, however, the oxime reaction has considerable importance, for by its means a monosaccharide may be changed into another sugar conlittle
has but
365
This is done by first making the oxime of taining one less carbon atom. the sugar and then heating the latter with acetic anhydride; the resulting acetyl-nitrile derivative is then heated with an ammoniacal solution of silver oxide which splits off the acetic acid
acid groups with formation of a lower sugar (Wohl's* synthesis). reaction in its simplest phase is represented as follows:
and hydrocyanic The
CH OH
2
CH OH
2
CH OH
2
(CHOH) 4
(CHOH) 3
(CHOH) 3
HCN
HC NOH
:
CHOH
CHO
nitrile
i-N
d-Glucose-oxime
d-Gluconic acid
d-Arabinose
Hydrocyanic acid
The hexose sugar

d-arabinose.
thus converted into the pentose sugar d-glucose In the same manner d-arabinose can be converted into
is
the tetrose sugar d-erythrose.

to
all
The reducing sugars, similar Cyanhydrine Reaction of Sugars. and with react aldehydes ketones, hydrocyanic acid forming a
group of compounds known as cyanhydrines.
characteristic
CH OH
2
CH OH
2
(CHOH) 4
HCN
(CHOH) 4
C O
:
CHOH
Hydrocyanic acid
d-Glucose-cyanhydrine (d-glucoheptonic acid
nitrile)
d-Glucose
reaction, as that of the oximes, while having but value in sugar analysis, has very great importance in sugar synthesis for by its means a monosaccharide may be 'built up into another sugar
little
The Cyanhydrine
having one more carbon atom.
This
is
done by
first
making the
cyanhydrine, saponifying this to form the corresponding acid, and then reducing the latter with sodium amalgam which produces the corres-
ponding sugar. The formation of glucoheptose from glucose as an illustration of this type of reaction.
is
given
CH OH
2
CH OH
2
(CHOH) 6
3H/)
(CHOH) 6
NH OH
4
C=N
d-Glucose-cyanhydrine
COOH
d-Glucoheptonic acid
Ammonia
CH OH
2
CH OH
2
(CHOH) 5
+
*
(CHOH) 5
HO
2
COOH
d-Glucoheptonic acid (lactone)
HC O
:
Hydrogen
d-Glucoheptose
Water
Ber., 26, 730.
366
SUGAR ANALYSIS
tose,
In the same manner, starting from the hexoses, mannose and galacmannoheptose and galaheptose can be derived. The heptoses by
the same cyanhydrine synthesis have been built up into the corresponding octoses CsHieOg and the latter in turn into the corresponding nonoses
CgHisOg.
*
For
details as to this
method
of forming sugars the
work
of
Nearly all reducing sugars, with exthe react at of ketoses, moderately warm temperatures with ception urea in presence of dilute sulphuric or hydrochloric acid to form a
group of compounds called ureides. The reaction is analogous to that with phenylhydrazine, the hydrogen of the amino group withdrawing the oxygen from the aldehyde group of the sugar. The reaction with
glucose
Fischer should be consulted. Ureide Reaction of Sugars.
and urea
2
is
given by
way
of example.
CH OH
(CHOH) 4
CH OH
2
(CHOH) 4
HC:0 +
Glucose
H N-CO-NH
2
HC:N-CO-NH
Glucose-ureide
Urea
HO + Water
2
The ureides are partly crystalline and partly amorphous bodies. In aqueous solution they are decomposed upon heating with evolution of
ammonia and
Semicarbazone Reaction
liberation of the free sugar. of Sugars.
Very similar to the reaction
of sugars with urea is that with semicarbazide; the latter in alcoholic solution combines with the aldoses to form a group of substances called
semicarbazones.
The
reaction with glucose
is
given as illustration.
2
CH OH
2
CH OH
(CHOH) 4
HC:O
Glucose
H N-N-CONH
2
(CHOH) 4
2
HC N - N - CONH
I
+ HO
2
Semicarbazide
Glucose-semicarbazone
Water
The semicarbazones are well-defined crystalline compounds; when warmed with benzaldehyde in alcohol solution they are decomposed into
with formation of benzaldehyde semicarbazone. Thiosemicarbazone Reaction of Sugars. Exactly similar to the reaction is the behavior of aldose with thiosemicarbazide. previous sugars
free sugar
The
reaction with glucose proceeds as follows:
CH OH
2
(
CH OH
2 4
CHOH)
HC:O + H N-N-CSNH
2
H
*
(CHOH)
2
Glucose
Thiosemicarbazide
HC:N-N-CSNH + H O Water Glucose-thiosemicarbazone

2
2
Ann., 270, 64; 288, 139.

The thiosemicarbazones
lar in
367
simi-
are well-defined crystalline
compounds
many
properties to the semicarbazones.
Reactions of Sugars with Aromatic Amines. The ease with which reducing sugars unite with compounds containing an amino
group, as
shown
in the case of the hydrazones, oximes, ureides, semi-
carbazones,
etc., is
further exemplified
by the
reactions of sugars with
different aromatic amines, such as aniline, toluidine, etc. Glucose, for example, reacts with aniline in alcoholic solution as follows:
CH OH
2
CH OH
2
(HCOH) 4
(HCOH) 4
2 6 5
H-C:0 + H NC H Glucose
Aniline
H-C:N-C H
6
Glucose anilide
Water
Reactions of Sugars with
Alcohols.
By
leading dry hydro-
chloric-acid gas into the solution of a reducing sugar in an alcohol the corresponding alcohol derivative of the sugar is formed. The com-
pounds thus prepared are called glucosides from their resemblance to the group of plant substances known under this name. The reaction of glucose with methyl alcohol is given as illustration.
7
/3
CH OH CHOH CHO FH""

2
CH OH
2
CHOH CH CHOH
CHOH HOH
:
/SCHOHJ)
a.
H-C
!O
+ H OCH
Methyl
H-C=0~--CH + H
3
Glucose
alcohol
Methyl
glucoside
Water
In the same manner glucosides of the other sugars have been made as methyl arabinoside, methyl xyloside, methyl rhamnoside, methyl The comfructoside, also of the other alcohols as ethyl glucoside, etc.
pounds thus prepared are well-defined crystalline substances, easily soluble in water, do not reduce Fehling's solution and do not react with
phenylhy drazine
.
reactions of the reducing sugars with alcohols are but little The synthetic glucosides have, used as a means of identification.
The
however, a great interest for the sugar chemist in other ways. Nearly all reducing sugars, exMercaptal Reaction of Sugars. react the with cept ketoses, mercaptans in presence of concentrated
hydrochloric acid to form mercaptals.
The
reaction with glucose and
ethyl-mercaptan
is
given as illustration.
368
SUGAR ANALYSIS
CH OH
2
CH OH
2
(CHOH) 4
(CHOH) 4
H-C:O
Glucose
4-
H H-S-C - 2
5 5
TT
H i/S-C 2
2
\S
CH
s 6
Ethyl-mercaptan
LGlucose-mercaptal
Water
The mercaptals
of the sugars are well-defined crystalline
soluble in hot water; they
compounds, do not reduce Fehling's solution and do not
react with phenylhydrazine. Reactions of Sugars with
sugars react with a large
The simple reducing Aldehydes. of aldehydes (formaldehyde, acetaldehyde, benzaldehyde, salicylaldehyde, furfural, etc.) to form a variety
number
The latter, for the most part, are of a of condensation products. gummy or sirupy nature and do not crystallize readily. The combination of glucose with acetaldehyde is given as an illustration of this
type of reaction.
CH OH
2
CH OH
2
(CHOH) 4
(CHOH) 4
H-C:O
Glucose
M O:C-CH
H-C(
XK u - CH > C
Acetaldehyde
Glucose-acetaldehyde
Reactions of Sugars with Polyvalent Phenols. The simple reducing sugars unite with different polyvalent phenols (resorcin, orcin, hydroquinone, phloroglucin, pyrogallol, etc.) to form a series of amorill-defined condensation products. The reaction is carried out in the cold in presence of hydrochloric acid. The following combination of arabinose with resorcin is given as an illustration of this type of
phous
reaction.
CsHioOs
Arabinose
-j-
CeHeC^
Resorcin
CnHwOe
Arabinose-resorcin
~h
HÔ.
Water
The condensation products of the sugars with polyvalent phenols when heated with concentrated hydrochloric acid are decomposed, showing the color
(see p. 341).
and
spectral reactions characteristic for each class of sugar
Reactions of Sugars with Acid Radicals.
In the
many
different
reactions previously described the aldehyde or ketone group of the sugar molecule is the one mostly involved. In the reactions of sugars
with acid radicals, as acetic and benzoic, the alcohol groups of the molecule are affected; the aldehydic characteristics of the sugar are also The number of acid deusually modified in the higher derivatives.
rivatives obtainable with a sugar
is
dependent upon the number of
369
In the case of hexoses having five such groups there alcohol groups. are mono-, di-, tri-, tetra- and penta- acetates and benzoates; with sugars of fewer alcohol groups the number of these combinations is
correspondingly less. Reaction of Sugars with Acetic Anhydride. are formed by heating with acetic anhydride.
Acetates of the sugars A mixture of different
acetates usually results during the reaction, the separation of these being effected by fractional crystallization or by the use of different
To obtain the highest acetates, the reaction must be carried out in presence of zinc chloride or some other condensing agent. The formation of glucose pentacetate is given as illustration of this type of
solvents.
reaction
CH OH
2
CH - CO
3
CH OCOCH
2
I
(CHOH) 4
+5
3
\ O
HC:O
Glucose
CH -CO
Acetic anhydride
(CHOCOCH HC O
:
3) 4
5CH COOH
3
Glucose-pen tacetate
Acetic acid
acetates of the sugars are amorphous, easily soluble subthe higher acetates are crystalline and less soluble in water. stances; with alcoholic potassium or sodium hydroxide, the acetates By warming
The lower
The lower aceare all easily saponified with regeneration of the sugar. tates of the sugars are copper reducing and exhibit other aldehydic properties; the higher acetates, as glucose-pentacetate, lack, however, many
aldehyde characteristics, such as formation of hydrazones and oximes. This is probably due to a stable lactonic rearrangement of the molecule * for glucose as shown by the following formula of Erwig and Konigs
pentacetate.
,CHOCOCH
/CHOCOC] CHOCOCH
HOCOCH
Reaction of Sugars with Benzoyl Chloride.
The
acetates of the
sugars owing to their solubility are not well adapted for the identification of sugars; the sugar benzoates, however, are marked by a high insolubility in water and their formation is sometimes used as a qualitative test for sugars.
*
Ber., 22, 1464, 2209.
370
SUGAR ANALYSIS
test,
according to the method of Baumann,* is carried out by of the sugar with benzoyl chloride in presence of a solution treating of the hydroxyl benzoic radical displaces the the sodium hydroxide; water. A number of of sodium chloride and formation with groups benzoates are usually formed in the reaction. In the case of glucose-
The
pentabenzoate the formation proceeds as follows:
CH OH
2
(CHOH) 4 + 5 C 6H COC1
5
+ 5 NaOH
Sodium hydroxide
is
CHO
Glucose
CH OCOC H (CHOCOC H )4 + 5 NaCl + 5 H O CHO

2
Benzoyl chloride
Glucose-pentabenzoate
Salt
Water
The Baumann
reaction
sufficiently delicate to detect 1 to 2
mgs.
glucose in 100 c.c. of water and is sometimes employed for testing urine; 100 c.c. of solution are well shaken with 2 c.c. of benzoyl chloride.
SPECIAL TESTS FOR REDUCING SUGARS

the second class of reactions for examining sugars belong the special tests pertaining to group identification; the reactions chosen for description may be divided for convenience into three general classes.
I.
To
II.
Analysis of hydrazones and osazones. Separation of products obtained by decomposition with concentrated hydrochloric acid.
III.
Color reactions with phenols in presence of concentrated mineral

acids.
I.
ANALYSIS OF HYDRAZONES AND OSAZONES AS A MEANS OF IDENTIFYING
SUGAR GROUPS
If
condition,
the hydrazone or osazone of a sugar has been separated in a pure an elementary analysis of the compound will serve to identify
the group to which the sugar belongs. The osazones, owing to their greater insolubility and ease of preparation, are best adapted for this
The determinations necessary for the identification of an osazone are those of the elements nitrogen and carbon; a determination of hydrogen is also usually included since this element can be
purpose.
determined with
determination.
little
extra trouble at the
same time
as the carbon
The elementary analysis of osazones and hydrazones is carried out by burning about 0.2 gin. of the substance over cupric oxide in a com*
Ber., 19, 3220.

bustion tube.
371
For nitrogen the combustion is carried out by Dumas 's carbon dioxide after complete displacement of The evolved nitrogen is received in a eudiometer over strong the air. potassium hydroxide solution and its volume measured. From the vol-
method
in a current of
calculated, making the necessary and temperature. For carbon and hydrogen the combustion is carried out by Liebig's method in a current of air or oxygen which must be perfectly dry and free from carbon dioxide. The evolved water is collected in weighed tubes, or spirals, containing concentrated sulphuric acid, and the evolved
is
ume
of gas the
weight of nitrogen
corrections for atmospheric pressure
carbon dioxide absorbed in weighed Liebig bulbs containing concentrated potassium hydroxide solution, or in U-tubes filled with soda lime (NaOH CaO). From the weights of water and carbon dioxide obtained the percentages of carbon and hydrogen are calculated. The percentage of oxygen in osazones and hydrazones is determined by subtracting the
sum
of the percentages of the other elements from 100. In the elementary analysis of osazones and hydrazones, as of all other nitrogen compounds, a spiral of copper should be placed in the combustion tube at the exit end in order to effect the reduction of
oxides of nitrogen. For complete details as to methods of combustion the chemist is referred to the standard textbooks upon organic analysis.
Having determined the elementary composition of an osazone or hydrazone, reference to a table of percentage composition will usually locate the class of sugar to which the compound belongs. In the following table the formula and percentage composition of phenylosazones are given for various groups of sugars.
Phenylosazone.
372
SUGAR ANALYSIS
SEPARATION OF PRODUCTS OBTAINED BY DECOMPOSITION WITH CONCENTRATED HYDROCHLORIC ACID AS A MEANS OF IDENTIFYING SUGAR GROUPS
II.
While an elementary analysis of osazones is one of the best means which a sugar belongs there are a number The most of other special group reactions which are of great value. and identification of charthe some these is of separation important acteristic decomposition product obtained by treating the sugar with
of determining the class to
concentrated sulphuric or hydrochloric acid. The latter acid drastic in its action and is the one most commonly used.
is
less
humus subthe decomposition products etc. obtained upon heating sugars with stances, aldehydes, acids, It is concentrated hydrochloric acid has already been mentioned.
The varied nature
of
found, however, that
when
this
treatment
is
one characteristic decomposition product

:
will
carefully controlled some predominate for each
The following equations, representing ideal particular group of sugar. are illustrations as of reaction, given types
I.
II.
III.
H CH Pentose C H (CH )0
C
6
12
= =
5
CH
5 5
Hexose
5
10
Levulinic acid
CHO
4
Furfural
(CH3 )02
Methylpentose
Methylfurfural
HO + Water + HCOOH Formic O + 3H Water HO + 3Water

2
acid
The above types of reaction hold true not only of the simple sugars above named, but also of the higher saccharides which yield these In fact the initial phase of the reaction in sugars upon hydrolysis.
case of the polysaccharides (sucrose, maltose, lactose, raffinose, starch,
pentosans, methylpentosans, etc.),

indicated.
is
purely hydrolytic, the simple
sugars formed being subsequently decomposed after the manner just
which
This reaction, Hexose Groups. due to Tollens * and has been extensively studied by his coworkers, has been employed with great success in detecting hexose groups in a large variety of plant and animal substances (cellular tissues of plants, nucleic acids of animal origin, etc.) Owing to the much
Levulinic Acid Reaction for
is
greater predominance of hexose-producing substances in nature the levulinic acid reaction is usually among the first tests applied in investigating materials of unknown composition.
Description of Test.
*
In carrying out the reaction 5 to 10 gms. of
Ann., 206, 207, 226; 243, 314; Ber., 33, 1286.

material are treated with 20 to 50
c.c.
373
of hydrochloric acid of 1.09 to
1.10 sp. gr. (18 to 20 per cent) in a flask provided with a rubber stopper and condensing tube, and heated in a boiling-water bath for 5 to 20
hours.
precipitate of
The brownish-colored liquid is then cooled and filtered from the humus substances; the filtrate is shaken out in a sep-
aratory funnel four times with ether, and the ether extract, after pouring through a dry filter, evaporated. The sirupy residue is then gently heated in an open dish to expel the formic acid (see previous equation I). If levulinic acid is present a drop of the sirup dissolved in water in presence of sodium carbonate and iodine will give a precipitate of iodoform, which can also be recognized by its characteristic odor.
The main portion

charcoal, filtered
of the sirup
is
dissolved in water, boiled with an
excess of zinc oxide (ZnO),
and then,
and evaporated.
The
after decolorizing with animal zinc salt of levulinic acid will
soon crystallize;
alcohol
is
and
ether,
the crystals are filtered off, washed with absolute and then converted into the silver compound. This
done by dissolving the zinc salt in 5 to 10 c.c. of water, adding silver nitrate slightly in excess of the equivalent amount and heating nearly to boiling, with addition of a little water until the precipitated silver
salt
has completely dissolved.

filtered.
little
animal charcoal
is
then added
and the solution

crystallizes will
show
levulinate of silver, CsHyOsAg, which under the microscope, in case the compound is
The
pure, hexagonal crystals or plates; if the crystals will be feather-like in appearance.

off,
compound is less pure the The silver salt is filtered
washed with cold water, pressed between filter paper and dried in a dark place over concentrated sulphuric acid. The per cent of silver in the salt is determined by strongly igniting a weighed portion in a porcelain crucible.
The
theoretical
amount
is
48.39 per cent Ag.
treating hexose sugars with hydrochloric acid will vary greatly according to the time of heating and other conditions of the experiment. Conrad and Guthzeit * obtained
yield of levulinic acid obtained
The
by
upon heating
c.c.
of acid (containing 4.87 gms.
10.5 gms. each of fructose, glucose, and galactose, with 50 HC1 gas) for 17 hours the following yield
of products.
Sugar.
374
SUGAR ANALYSIS
these results
it
From
appears that of the three hexose sugars fruc-
tose gives the largest yield of levulinic acid and galactose the least. That this is due largely to the greater resistance of glucose and galactose toward the acid was shown by the fact that at the end of the above
experiments considerable quantities of these sugars were still undecomposed (in case of glucose 26 per cent). The yield of levulinic acid
method to be of any quantitative value. Furfural Reaction for Pentose Groups. This reaction, which is also due to Tollens,* has been of the greatest value not only as a means of detecting the presence of pentose carbohydrates but also as a means of their quantitative estimation.
is
too variable for the
The reaction of the pentose sugars with hydrochloric acid proceeds much more nearly according to the equation (II, p. 372) than the reaction of the hexoses, the formation of humus substances being correspondingly less. The following graphic equation shows the decomposition of a pentose sugar into furfural.
CH-CH-iOHi xOjH CH-C x V :l c:o J

I
CH:CH
........
......
+ in-p' CH C \C-0 M
'
3H2
JQH...H!
Pentose (150 parts)
Furfural (96 parts)
Water
(54 parts)
The theoretical yield of furfural, according to the above equation, 64 per cent; actual determinations of the furfural, obtained by distilling weighed amounts of the pentose sugars, arabinose and xylose, with hydrochloric acid, give about 47 per cent in case of arabinose and
is
about 57 per cent in case of xylose and 90 per cent respectively of the
yields
which are about 75 per cent
theoretical.
In carrying out the qualitative test about Description of Test. 5 gms. of substance are heated in a distillation flask with 100 c.c. of hydrochloric acid of 1.06 sp. gr. and successive portions of about 30 c.c.
a receiver, new portions of acid being added to the flask for each quantity distilled. The distillates are then tested for the presence of furfural; the latter in large amounts can usually be detected by its
distilled into
pleasant aromatic odor
somewhat resembling that
of bitter
is
almond
oil.
The presence
agent
is
of very small amounts of furfural Schiff's reaction with aniline or xylidine acetate.
best indicated
by
Aniline acetate re-
best prepared according to Tollens by mixing in a test tube equal volumes of aniline and water and then adding with constant shak*
Landw.
Vers.-Stat., 39, 425.

ing glacial acetic acid drop
clear.
375
by drop until the milky solution becomes prepared by moistening strips of filter paper with the aniline-acetate solution. Application of a drop of distillate containing furfural, even in minute traces, will cause the aniline-acetate
Test paper
is
paper to turn a bright cherry red.
The presence
first
of furfural in the distillate
may
also be indicated
by
neutralizing the acid solution with sodium carbonate and then addFurfural if presing a solution of phenylhydrazine acetate and stirring. ent is precipitated as furfural-phenylhydrazone, C 4 3 2 HC6H5,
H OCHN
which melts at 97 to 98 C.
better precipitating agent for furfural than phenylhydrazine is A solution of this compound in hydrochloric acid when phloroglucin.
added to a
distillate
containing furfural will cause an immediate dark-
ening of the solution with final precipitation of furfural-phloroglucide, according to equation:
C H4
5
CH
6
03
CnH O
8
HO
2
Furfural
Phloroglucin
Furfural-phloroglucide
Water
Limitations of Furfural Reaction for Pentoses. While all carbohydrates containing a pentose group yield large amounts of furfural upon distillation with hydrochloric acid, it must also be borne in mind that
other substances have the same property.
as starch, cellulose, sucrose, glucose, etc., give small
All hexose carbohydrates such amounts of furfural
upon distillation with hydrochloric acid but the

terfere seriously
yield
is
too small to in-
with the test for pentoses. Two substances, however, of a non-pentose nature are especially marked by their property of yielding furfural upon distilling with acids and hence require brief mention. These are glucuronic acid and oxycellulose. Glucuronic acid is an aldehyde-acid derivative of glucose and has
the formula
bacteria
it is
COH(CHOH) COOH. By
4
the action of putrefactive
converted into the pentose sugar 1-xylose.
H O
10
CH
5
10
C0
Glucuronic acid
Xylose
Carbon dioxide
is
The intimate relationship of glucuronic acid to the pentoses shown by the reaction upon distilling with hydrochloric acid.
also
CH O
6
10
Glucuronic acid
Furfural
O + C0 + 3H Carbon Water
2
dioxide
Glucuronic acid is sometimes found in the urine, especially after the ingestion of chloral, menthol, camphor, turpentine, acetanilide, alkaloids and many other compounds. Under such conditions a combination takes place in the animal organism between the ingested
compound
376
SUGAR ANALYSIS
and the glucuronic acid, the latter apparently being formed as an oxidation product of glycogen. The glucuronic-acid derivative, which is excreted in the urine, may be mistaken for a pentose sugar if the chemist relies solely upon such tests as the furfural reaction and reduction of
metallic salt solutions.
One means of determining the presence of glucuronic acid is by means of p-bromophenylhydrazine, which was found by Neuberg * to
The give a characteristic glucuronic-acid derivative, C^HnOr^Br. exact nature of the compound, whether hydrazone or hydrazide, was not determined. The solution to be tested is heated in a water bath at
60 C. with 5 gms. of p-bromophenylhydrazine chloride and 6 gms. of
sodium acetate.
filtered off
If glucuronic acid is present yellowish needle-like crys-
tals will separate in 5 to
10 minutes.
The
solution
is
cooled, the crystals
again heated as before; a second crop of thus be obtained which are filtered off again and the crystals may continued until no more The combined precipiprocess crystals form.
filtrate
and the
tates are thoroughly washed with warm water and then with absolute alcohol. Recrystallized from 60 per cent alcohol the crystals melt at
236 C. The crystals dissolved in a mixture of 6 c.c. absolute alcohol and 4 c.c. pyridine have a strong levorotation, [O\D = 369.
Spectroscopic methods for distinguishing between pentoses and glucuronic acid will be described under the color reactions for sugar groups.
when treated with different oxidizing agents, such as nitric chromic acid, hypochlorous acid and permanganate, undergoes a The oxycellulose derivatives formed under such partial oxidation.
Cellulose,
acid,
conditions have the property of yielding furfural
upon
distillation
with
hydrochloric acid.
According to the researches of Tollens and Faber

consist of mixtures of cellulose
f oxycelluloses
(CeHioOs^
6 )n.
in different porportions
with
amount of celloxin in the oxycellulose the greater the yield of furfural upon distillation with hydrochloric acid. Cotton, for example, upon treatment with
greater the
nitric acid at
an oxy-derivative celloxin (C 6 H 8
The
100 C. for different periods of time, gave the following
results
Time
of
treatment.

The
yet been isolated)
is
377
yield of furfural calculated to pure celloxin (which has not as is about 12 per cent.
The oxycelluloses are widely distributed in nature and if reliance based exclusively upon the furfural reaction erroneous conclusions may be formed as to the occurrence of pentose carbohydrates in plant
The oxycelluloses may be easily distinguished, however, from pentosans by the fact that they yield glucose exclusively upon hydrolysis with acids, the hydrolytic products giving none of the rematerials.
actions (osazone, color tests, etc.) characteristic of the pentoses. In the Methylfurfural Reactions for Methylpentose Groups.
same way that all substances containing pentose groups yield furfural upon distilling with hydrochloric acid, those materials containing methylpentose groups yield methylfurfural. ogous to that described upon page 374.
The
reaction
is
perfectly anal-
iOH H; >CH, -C-iOHi

| |
= y
-%
H
Water
(54 parts)
X C:O
jpH"H|H
Methylpentose (164 parts)
Methylfurfural (110 parts)
The theoretical yield of methylfurfural from methylpentose accordIn actual distillation exing to the above reaction is 67.07 per cent. periments with the methylpentoses, fucose and rhamnose, only from
35 to 40 per cent methylfurfural
theoretical
is
obtained or 50 to 60 per cent of the
amount.
In testing natural products for the presence of methylpentose groups, the material is distilled with hydrochloric acid of 1.06 sp. gr. in exactly the same manner as described for pentoses and the distillate
If no furfural is present in the distillate the presence of methylfurfural will be indicated by aniline-acetate paper, which in this instance is colored yellow. If pentosans are also present
tested for methylfurfural.
in the plant material being examined, as is nearly always the case, the presence of furfural in the distillate will color the aniline-acetate paper
red and completely

action.
mask the yellow
color of the methylfurfural re-
Other tests must, therefore, be employed to detect the presence

*
of methylfurfural.
has devised a reaction by which 1 part methylfurcan be detected in presence of 9 parts furfural. A small amount of the solution to be tested is added to a mixture containing 3 volumes
Maquenne
fural
Compt.
rend., 109, 573.
378
SUGAR ANALYSIS
95 per cent alcohol and 1 volume concentrated sulphuric acid and the whole gently warmed. The development of a bright grass-green color throughout the body of the solution indicates the presence of methylfurfural.
Spectral reactions for methylfurfural will be described in a succeed-
ing section.
Reactions for Tetrose and Triose Groups. Excepting the hexoses, but few and experiments have been made methylpentoses, pentoses other reactions of the sugar groups with hydrochloric acid. concerning
* show that 1-erythrose Experiments of Tollens and Ellett with acid into lactic acid. heating hydrochloric composed upon
is
de-
The
reaction
may
proceed as follows:
CH
4
CH
3
+
may
is
CH
Tetrose
Lactic acid
Formaldehyde
Tollens and Ellett suggest that the above

for tetrose groups, just as levulinic acid
fural
be a general reaction
fur-
formed from hexoses, from pentoses, and methylfurfural from methylpentoses.

of considerable methylglyoxal
The formation
CH
CO COH
by
heating dioxyacetone, CaHeOa, with sulphuric acid has been observed by Pinkus.f This may perhaps be a group reaction of trioses.
Further investigations require to be made upon the tetroses and any results from the above observations can be applied to sugar analysis.
trioses before
III.
COLOR AND SPECTRAL REACTIONS AS A MEANS OF IDENTIFYING SUGARS

study of the color reactions and absorption spectra which solu-
tions of different sugars give with various phenols as a-naphthol, orcin, resorcin, naphthoresorcin and phloroglucin, in presence of concentrated
sulphuric or hydrochloric acids offers frequently a most rapid as well as most reliable method for detecting sugar groups. Color Reactions of Ketoses. Reference has already been made
(p.
tion
340) to the greater ease with which solutions of ketoses show coloraphenomena in contact with concentrated sulphuric acid. The same
been noted with the colorations produced with sugars and a-naphthol and sulphuric acid, and this has been utilized as one means
fact has
of detecting the presence of ketose sugars in mixtures.
a-Naphthol Test.
Pinoff { has modified the a-naphthol test for sugars
of 750 c.c. 96 per cent alcohol and 200 gms. concentrated sulphuric acid as the condensing agent. By treating in a test
by using a mixture
*
Ber., 38, 499.
f Ber., 31, 31.
t Ber., 38,
3314.

tube 0.05 gm. of sugar with 10
c.c. c.c.
379
of the alcohol-acid
mixture and
of alcoholic a-naphthol (5 gms. a-naphthol dissolved in 100 c.c. 0.2 cent 96 per alcohol) and heating in boiling water, Pinoff obtained red colorations which in case of sugars containing ketone groups appeared
almost immediately; with the aldose sugars 20 minutes or more elapsed The following table for 1 1 different sugars before coloration developed. of time Pinoff the heating before coloration, the number of gives by bands shown by the solution before the spectroscope and absorption
the position of the bands with reference to the absorbed.
wave length
of the light
TABLE LXVIII
Giving Absorption Spectra of Sugars with a-Naphthol and Sulphuric Acid in Alcohol
Sugar.
380
SUGAR ANALYSIS
While diluting the acid-alcohol mixture has practically eliminated

the aldoses from the reaction, it has also materially lessened the sensibility of the test for the ketoses.
The most convenient color test for distinguishing Resorcin Test. ketose from aldose sugars is the color reaction with resorcin and hydro* test. The test was chloric acid generally known as Seliwanoff's
originally regarded as peculiar to fructose,
but later experiments have
given by sorbose, tagatose, the keto-pentoses and all other sugars having a ketone group. The reaction is carried out by mixing in a test tube 10 c.c. of the
shown that
it is
be tested with 10 c.c. of 25 per cent hydrochloric little resorcin (about the tip of a knifebladeful), and a acid, then adding If fructose or other ketose is present flame. a small over heating gently will color eosin-red a fiery develop, which upon cooling and standing will deposit as an amorphous powder mixed with humus decomposition
clarified solution to
products. If the acid solution
is
made
alkaline with soda
and then shaken with
amyl alcohol, the red coloring matter is dissolved with a greenish If a few drops of absolute alcohol be now added the fluorescence.
color
If
becomes a beautiful rose
red.
the red-colored solutions obtained
by
Seliwanoff's reaction be ex-
amined before the spectroscope a

in the blue near the
It is
distinct absorption
band
will
be noted
line.
(See Fig. 165.)
important in making the test with resorcin that an excess of hydrochloric acid be avoided. The percentage of acid in the final mixIf too much strong acid is present, ture should be about 12 J per cent. with resorcin and form pinkand other aldoses will react also glucose colored solutions; the latter, while lacking the intensity of color obtained with the ketoses, may nevertheless lead to erroneous conclusions. The resorcin reaction obtained with glucose may be due to a slight transformation of this sugar into fructose. Ost, as a matter of fact, has succeeded in effecting such a transformation by treating glucose in the cold with strong sulphuric acid.
Pinoff f has modified the resorcin test for ketoses
by using the
alcohol-sulphuric-acid mixture previously described as the condensing In making the test 0.05 gm. of sugar was treated in a test agent.
tube with 5
c.c.
0.2 c.c. of a 5 per cent resorcin solution
of the alcohol-sulphuric-acid reagent, 5 c.c. alcohol and and the mixture placed in boil-
The following table for 11 different sugars by Pinoff gives ing water. the length of time required for development of color, the number of
*
Ber., 20, 181.
Ber., 38, 3314.

absorption bands and the position of the bands with reference to length of light absorbed.
381
wave
TABLE
LXIX
Giving Absorption Spectra of Sugars with Resorcin and Sulphuric Acid in Alcohol
Sugar.
382
SUGAR ANALYSIS
pentosans, gave, upon heating with an equal volume of concentrated hydrochloric acid and a little phloroglucin, a beautiful violet-red color. The colored solution thus obtained when viewed before the spectro* scope was found by Tollens and Allen to show a sharp black absorpof the and E lines. in the tion band spectrum between the yellow The violet-red solution obtained in the phloroglucin reaction for
pentoses soon becomes turbid with deposition of a dark-colored precipiIf the turbid solution is allowed to stand 3 to 5 minutes, then tate.
cooled, filtered
rapid solution
filter
is
and the precipitate washed with cold water on a small and then dissolved in 95 per cent alcohol, a permanent red obtained which is perfectly adapted to the study of ab-
sorption spectra.
If the color is too deep it can be reduced by careful " " absatz dilution with 96 per cent alcohol. (Tollens's method.) The same color reaction of the pentoses with phloroglucin and
hydrochloric acid
not
given by glucuronic acid and its derivatives, but by oxycellulose. The test, therefore, while enabling the chemist to distinguish between such furfural-yielding substances as pentosans and
is
oxycellulose, does not permit the distinction and pentoses (as for example in urine).
between glucuronic acid
If the reaction for the pentoses just described be Orcin Test. carried out with orcin in place of phloroglucin a violet-blue coloration The solution, however, becomes rapidly turbid with deis obtained.
If the latter is filtered position of a bluish-colored flaky precipitate. " " off and dissolved in alcohol by Tollens's absatz method a blueis obtained which shows before the spectroscope a very dark band almost exactly over the D line of the spectrum. The sharp same reaction is also obtained with glucuronic acid. Bial f has made the orcin reaction more sensitive by carrying out
colored solution
the test in presence of a little ferric chloride. In this manner it found possible to distinguish between pentoses and glucuronic acid.
is
Bial's orcin reagent is prepared by dissolving 1 gm. orcin in 500 c.c. hydrochloric acid of 1.15 sp. gr. (30 per cent) to which 20 drops of
an
officinal solution of ferric chloride
(liquor ferri sesquichloridi) are
added.
In making the test 4 to 5 c.c. of the reagent are heated in a test tube to boiling; the solution is removed from the flame and a few drops If pentoses are (never over 1 c.c.) of the solution to be tested added.
present a vivid green color will develop almost immediately; the reaction is not given under the above conditions with glucuronic acid.
*
Ann., 260, 289. Biochem. Zeitschrift.,
3,
323.

also
383
BiaFs test has been studied and generally confirmed by Sachs,* and by Tollens and Lefevre.f The last-named authorities found that
a dilute solution of glucuronic acid produced no perceptible coloration under the conditions prescribed by Bial, but that if the solution was
heated for any length of time a green color speedily developed. The cause of the retardation is explained by the slower decomposition of glucuronic acid by hydrochloric acid as compared with the pentoses; such a difference in the rate of decomposition is also noted between the
pentose sugars themselves, xylose, for example, giving a coloration with Dial's reagent in a shorter time than arabinose.
The green solution obtained by BiaFs reaction shows before the spectroscope a dark absorption band in the red between the lines B and line C and a second band in the yellow covering the position of the of the spectrum.
Tollens and Naphthoresorcin Test for Pentoses and Glucuronic Acid. Rorive J have found that when solutions of different sugars are heated
little naphthoresorcin in presence of an equal volume of concentrated hydrochloric acid (1.19 sp. gr.) characteristic colored solutions and deposits are formed.
with a
With the pentoses arabinose and xylose a red color develops on heating followed by a bluish turbidity. The deposit dissolves in alcohol to a reddish-brown solution with beautiful green fluorescence, showing a weakly-defined absorption band in the green.
Glucuronic acid gives with naphthoresorcin and hydrochloric acid a bluish turbid solution with blue deposit. The alcoholic solution of
the latter
is
dark
absorption
a beautiful blue, only slightly fluorescent, and shows a line of the band in the yellow covering the
spectrum.
The naphthoresorcin
Tollens
in the following
test for glucuronic acid has been improved by way. The deposit of coloring matter is
treated with ether instead of alcohol; if glucuronic acid is present the ether is colored a violet blue and shows before the spectroscope an
absorption band in the yellow, its center lying a D sodium line (i.e., toward the green).
little
to the right of the
The naphthoresorcin deposits obtained with sugars (pentoses, hexoses, etc.) in presence of hydrochloric acid are insoluble in ether and so do not appear in the reaction. The presence of sugar and also
of foreign organic matter, as in urine,
may change
the color of the ether
solution from the violet blue characteristic of pure glucuronic acid to a

*
Biochem.
Zeitschrift., 1, 384.
{ Ber., 41,
1783.
t Ber., 40, 4520.
Ber., 41, 1788.
384
violet, red, or
SUGAR ANALYSIS
reddish brown.
The
the yellow part of the

B C
spectrum
will not,
characteristic absorption band in however, be interfered with.
Indigo
Violet
Fructose,
resorcin
and
hydro-
chloric acid.
Sucrose, a-naphthol
acid.
and sulphuric
Arabinose, phloroglucin and hydrochloric acid.
Methylfurfural, phloroglucin and
hydrochloric acid.
Methylfurfural and hydrochloric

acid.
Fig. 165.
Absorption spectra given by different sugars.

test as prescribed by Tollens is made as folthe solution (urine, etc.) to be tested are treated in
The naphthoresorcin
lows: 5 to 6
c.c. of

a 16
385
mm. wide test tube with \ to 1 c.c. of a 1 per cent solution of naphthoresorcin in alcohol and an equal volume of hydrochloric acid The solution is carefully heated to boiling and of 1.19 sp. gr. added.
minute over a small flame. The dark-colored solution minutes and then cooled under a stream of cold water; an equal volume of ether is then added and the whole thoroughly
then kept for
is
set aside for 4
After the acid solution has settled the ether layer will be colored blue or bluish violet to red, in case glucuronic acid is present, and, if the tube is held before the spectroscope, will show the character-
shaken.
In case the ether does not sepaline. istic absorption band near the If the rate readily a drop or two of alcohol will hasten the process. ether solution is too deeply colored for spectroscopic examination more ether is added until the color is reduced and the unabsorbed part of the
spectrum made
visible.
deposits of the pentoses and other sugars being insoluble in ether separate as a layer between the colored ether and the lower acid solution.
The naphthoresorcin
The color reactions for Color Reactions of Methylpentoses. detection of methylpentoses may be divided into two classes: (1) color
the distillate obtained by distilling methylpentoses with or methylpentosans hydrochloric acid; (2) color reactions made these substances without distillation. The color reactions directly upon
reactions
of the first class are in reality
made upon
color
reactions
which reference has already been made.
It remains,
of methylfurfural to however, to describe
some
of the spectral reactions of methylfurfural.
Tollens and Widtsoe * have Spectral Reactions of Methylfurfural. detected the presence of methylfurfural in the hydrochloric acid distillate from various plant materials by mixing a few cubic centimeters
of the solution
and gently warming.

present.
with an equal volume of concentrated hydrochloric acid If the solution is colored yellow methylfurfural is The yellow solution viewed before the spectroscope will show
a dark absorption band between the green and blue of the spectrum near the F line. If much methylfurfural is present the band will gradually darken and broaden, the increase in width extending toward the
violet
furfural the violet
green,
and leaving the green unaffected. With considerable methylend of the spectrum is completely extinguished, the however, always remaining clear and transparent. Furfural does
it
not give this reaction although

present in large amount.
may
affect the delicacy of the test

1
if
The
*
reaction, however, will indicate
part of
methylfurfural in presence of 64 parts furfural (^V drop methylfurfural

Ber., 33, 146.
386
SUGAR ANALYSIS
By
use
in presence of 2 drops furfural in 10 c.c. of hydrochloric acid).
of this test Tollens in different
and Widtsoe were able to detect methylpentosans gums, sea weed, leaves of different kinds of trees and a
large variety of other plant substances.
and Oshima * have rendered the spectral reaction for methylfurfural more sensitive by carrying out the test in presence of
Tollens
phloroglucin; 5 c.c. of the hydrochloric acid distillate are treated with 5 c.c. of concentrated hydrochloric acid and a few cubic centimeters of
a solution of phloroglucin (in hydrochloric acid of 1.06 sp. gr.) added. After 5 minutes the solution is filtered from the greenish-black precipitate of furfural phloroglucide;
is if
the
filtrate is
colored yellow or reddish
yellow methylfurfural present. scope a dark absorption band in the blue.
The
solution gives before the spectro-
On
long standing the solu-
tion deposits a red precipitate of methylfurfural phloroglucide which is readily distinguished from the dark-green furfural compound. Ab-
sorption spectra of methylfurfural are shown in Fig. 165. The vivid color reactions of the pentoses with orcin and phloroglucin are not obtained with the methylpentoses. Naphthoresorcin, however,
was found by Tollens and Rorive to give a deposit of coloring substance with the methylpentoses, rhamnose and fucose, when heated in
presence of hydrochloric acid. The alcoholic solution of the deposits showed a violet blue color with an exceedingly brilliant green fluorescence, which showed before the spectroscope an absorption band in the
line and a second band in the green. yellow over the There are a number of other color spectral reactions which have not
been described; these belong, however, more to the reactions of individual sugars and will be given under the description of these. A few characteristic absorption spectra, useful in testing for sugars,
are
shown
in Fig. 165.
Reactions of the Non-reducing Sugars

of sugars, which do not reduce Fehling's solution, all belong to the higher di-, tri- and tetrasaccharides and include sucrose, trehalose, raffinose, melezitose, gentianose, lactosinose, secalose, lupeose
The comparatively small number
and stachyose.
The
soluble polysaccharides,
such as dextrin, inulin, glycogen, etc., although not classified as sugars, are sometimes included for convenience in the group of non-reducing
saccharides.
free aldehyde, or
ketone group, to which the reducing sugars owe

*
their peculiar reactivity in the formation of hydrazones, oximes, ureides,

Ber., 34, 1425.
387
mercaptals, etc., is lacking in the non-reducing sugars, and the inability of the latter to reduce Fehling's solution, or to react with phe-
nylhydrazine, dilute alkalies, hydroxylamine, etc., is thus explained. The non-reducing sugars give many of the color and spectral reactions of the reducing sugars, sucrose and raffinose, for example, giving the a-naphthol reaction with sulphuric acid and Seliwanoff s reaction
with resorcin and hydrochloric acid. But as previously explained these reactions are not given by the original non-reducing sugar, but by the
reducing sugars derived from this by the hydrolytic action of the acid used in making the test.
carefully controlled hydrolysis
by means
of acids or
enzymes,
combined with quantitative measurements of changes in polarization or in copper-reducing power, is the most reliable test for the presence of non-reducing sugars. Methods involving this principle have been described under the inversion methods for determining sucrose and raffinose, and other examples will be given under quantitative chemical methods. Individual tests will be described under the heading of each single sugar in Part II of this Handbook.
CHAPTER XIV
REDUCTION METHODS FOR DETERMINING SUGARS
THE
principal chemical
methods
for determining sugars are based
upon the property which

glucose, lactose
all
aldehydes and ketones have of reducing
alkaline solutions of certain metallic salts.
The reducing
action of
and other sugars upon alkaline solutions of copper, and other metals has already been mentioned. bismuth silver, mercury, In the case of silver and glucose, for example, the reaction when carefully controlled proceeds as follows:
CH O
6 12
Glucose
Silver oxide
Ag2p
= 18 Ag +
Silver
(COOH) 2
Oxalic acid
H 0. + 3 Water.
2
If the
amount
weight of reduced silver be determined for this reaction, the But unfortunately the reof glucose can easily be estimated.
ducing action of sugars upon metallic salts does not proceed with the quantitative precision of the above equation; the reduction is rarely complete and the amount of reduced metal varies with the conditions
The latter difficulty is obviated, however, in practhe by controlling process so that the same weight of reduced metal is always obtained for the same weight of sugar.
of the experiment.
tice
Of the various alkaline solutions of metals those of copper are employed almost exclusively in sugar analysis.
COPPER REDUCTION METHODS

Early Methods.
salts of
copper has been
The reducing known since
action of sugars the

first
upon
different
beginning of chemistry.
Trommer,*
solution as a
in 1841, first noted the value of alkaline copper-sulphate means of distinguishing grape from cane sugar. Tromin
mer 's method was improved
1844 by Barreswil
who made
the im-
portant discovery that addition of potassium tartrate to the alkaline Barreswil's copper-sulphate solution greatly increased its stability.
method was volumetric; the sugar solution was slowly added to the boiling copper reagent, which had previously been standardized against pure glucose, until the blue color was just discharged.
*
Ann., 39, 360.
t Journal
de Pharmacie 388
[3], 6,
301.

Fehling's Method.
of the alkaline copper
389
Pehling,* in 1848, first worked out the details method, as they now stand, and the copper-sul-
phate and alkaline-tartrate reagent has since been called by his name. The copper solution employed by Fehling consisted of 40.00 gms. copper sulphate, CuS0 4 .5 2 O, 160 gms. neutral potassium tartrate and
600-700 gms. sodium hydroxide sol. of 1.12 sp. gr. dissolved in water to This is equivalent to 34.65 gms. CuS0 4 .5 H2 1154.4 c.c. dissolved to 1000 c.c., the proportion used by nearly all subsequent workers down
to the present time. Fehling's solution contains 8.822 gms. copper to 1000 c.c. or 0.008822 gm. to 1 c.c. According to Fehling's experiments 1 c.c. of his soluof anhydrous glucose, or 1 part In terms of the molecular weight reduced 1.765 parts copper. glucose of glucose the ratio would be 180 X 1.765 = 317.6. Dividing this value by 63.6, the atomic weight of copper, the atoms of copper reduced
tion
was exactly reduced by 0.005 gm.
by one molecule of glucose is found to be five. The reduction ratio 1 5 was regarded as constant by Fehling and was so employed by all chemists until Soxhlet f showed in 1878 that the ratio between sugar and amount of copper reduced was not a constant but varied according to the excess of copper which is present during the reaction. The more modern methods of sugar determination, which employ I. VoluFehling's solution, may be divided into two general classes. metric methods based upon the complete reduction of a measured volume of standard solution. II. Methods based upon a gravimetric
:
or volumetric determination of the reduced copper.
VOLUMETRIC METHODS BASED UPON THE COMPLETE REDUCTION OF A MEASURED VOLUME OF FEHLING'S SOLUTION
Soxhlet's Method.
Owing
to the decomposition which takes
place in the mixed copper-sulphate and alkaline-tartrate solution upon standing, the two solutions employed in the Soxhlet and all other
modern methods
sist
of the following:
CuSO4 .5 H2
The solutions conjust before using. Solution A, 34.639 gms. of pure crystallized Solution are dissolved in water and made up to 500 c.c.
are
mixed only
B, 173 gms. of Rochelle salts are dissolved in water, 100
c.c. of
a solu-
tion of caustic soda, containing 516 gms. per liter are added, and the volume completed to 500 c.c. Previous to analysis mix equal
NaOH
A and B. Before using the mixed copper reagent, it shoulc} be standardized against glucose, invert sugar, lactose, etc., according to the needs of * t J- prakt. Chem. [2], 21, 227. Ann., 72, 106; 106, 75.
volumes of solutions
390
SUGAR ANALYSIS
Since reducing sugar in sugar-cane, sugar-beet and most analysis. other food products is most usually expressed as invert sugar, the latter A standard solution of is most commonly used for standardization.
invert sugar has also an advantage in being easily prepared. Standard Invert Sugar Solution. Method of the Association of Dissolve 4.75 gms. of pure sucrose in Official Agricultural Chemists.*
75
c.c.
of water,
add 5
c.c.
of 38.8 per cent hydrochloric acid
aside during a period of 24 hours at a temperature not Neutralize the acid exactly with dilute sodium hydroxide
and set below 20 C. and make up
to 1000 c.c.; 100 c.c. of this solution contains 0.500 gm. of invert sugar. The amount of standard invert sugar solution necessary to reduce 100 c.c. of the mixed copper reagent is determined according to the
details described in the next paragraph.
Application
to
Analysis of Sugar Products.
Method
of the Associa-
a preliminary titration to determine the approximate percentage of reducing sugar in the material under examination. Prepare a solution which contains approxtion of Official Agricultural Chemists.^
Make
imately
per cent of reducing sugar.
Place in a beaker 100
c.c. of
the
mixed copper reagent and approximately the amount of the sugar Boil for two minutes. solution for its complete reduction. Filter through a folded filter and test a portion of the filtrate for copper by use of acetic acid and potassium ferrocyanide. Repeat the test, varying the volume of sugar solution, until two successive amounts are found which differ by 0.1 c.c., one giving complete reduction and the
other leaving a small
two readings
amount of copper in solution. The mean of these taken as the volume of the solution required for the complete precipitation of 100 c.c. of the copper reagent. Under these conditions 100 c.c. of standard copper reagent require
is
0.475 gm. of anhydrous glucose or 0.494 gm. of invert sugar for comCalculate the glucose by the following formula: plete reduction.
V =
W
rp,
the volume of the sugar solution required for the complete reduction of 100 c.c. of standard copper reagent. the weight of the sample in
1 c.c.
of the sugar solution.
I hen
100
_
r
or
100
-
X 0.475 = per cent of glucose, X W X 0.494 = per cent of invert sugar. /\ w

.
solution are placed

*
In making the test for unreduced copper a few drops of the filtered upon a white test plate, acidified with a few drops of
Bull. 107 (revised)
U.
S.
f Ibid.
391
10 per cent acetic acid and a drop of 2 per cent potassium-ferrocyanide brown coloration indicates the presence of unresolution added.
duced copper.
Volume of Fehling's Solution Reduced by Different Sugars. The ratio between volume of standard Fehling's solution and the amount
was de-
of different sugars, just sufficient to cause complete reduction, * termined by Soxhlet to be as follows:
TABLE
Volume
of Fehling's solution
LXX
different sugars.
reduced
by
392
SUGAR ANALYSIS
preliminary determination and then making up a fresh sugar solution so that the percentage of reducing sugar shall be 0.1 per cent, 0.5 per cent or 1.0 per cent, etc., according to the volume of Fehling's solution
taken and the individual preference of the chemist. In this manner approximately the same volume of sugar solution is always used for reducing the same volume of copper reagent, and under such conditions, with a uniform method of boiling, the most accurate results are
obtained.
power is also obtained whether the sugar added to the copper reagent in small portions, with successive periods of boiling, or only in one portion with one period of The most accurate results are secured where the test is made boiling. with the entire volume of sugar solution, necessary for complete reduction, with only one period of boiling. The following example will give an illustration of the application of the method:
difference in reducing
solution be
20 c.c. of Fehling's solution diluted with 80 c.c. of water were Example. found to require for reduction exactly 20.2 c.c. of standard invert sugar solution or 0.101 gm. 50 gms. of sugar-cane molasses were diluted to 1000 c.c. Of this solution about 8 c.c. were required to discharge the blue color of 20 c.c. Fehling's solution.
of the sugar solution (4 gms. molasses) were then made up to 200 c.c. 0.02 gm. molasses). Of this solution 19.6 c.c. when boiled with 20 c.c. Fehling's solution and 80 c.c. of water for 2 minutes showed incomplete reduction by the ferrocyanide test and 19.8 c.c. complete reduction. Calling 19.7 c.c. the volume of sugar solution necessary to reduce the
80
c.c.
(1
c.c.
20
c.c. of
Fehling's solution, then
^^ U.U.Z X
19.7
^=
25.64 per cent invert sugar in
the molasses.
The Ferrocyanide Test for Copper. Several methods are followed for making the ferrocyanide test for unreduced copper. It sometimes happens that the cuprous oxide is precipitated in a very finely
divided form, and gives annoyance by running through the filter. One method of making the test is to superimpose several small strips
of filter
paper and allow a few drops of the solution to fall upon the upper paper. The moistened area upon the second or third underlying strip is then treated with a drop of ferrocyanide solution acidified with
The appearance of a brown spot indicates the presence of unreduced copper. Another method of removing the portion of solution to be tested is by means of a Wiley or Knorr filtering tube, which is prepared as folacetic acid.
lows:

Wiley's Filter Tube.
393
The Wiley
filter
tube, Fig. 166a, consists of
a piece of glass tubing, 5 to 7 mm. in diameter and 20 to 25 cm. long, one end of which has been softened in a flame and
A piece then pressed out so as to form a shoulder. is then stretched tightly over the end and In using the tube the tied securely by a thread.
of fine linen
covered end
is
dipped into water containing in suspen-
sion finely divided asbestos, and a film of the latter spread over the surface of the filter by suction at the
upper end. A small portion of the liquid to be tested sucked into the tube and then poured from the open end onto the test plate. Knorr's f modification of the Wiley tube is of smaller diameter and contains a perforated platinum disk in place of the linen (Fig. The disk is coated with asbestos and the liquid 1666). withdrawn for testing as with the Wiley tube. The filter tubes should not be reused until after cleaning in dilute nitric acid and washing with water. A method due to Ross,| and Method of Ross.
is
in Louisiana, is to dip the point of a small folded filter, held by means of forceps, below the surface of the hot solution in the cas-
employed quite extensively
and withdraw a few drops of the clear liquid from the interior of the filter by means of a medicine The method is simple, and pardropper (Fig. 167). ticularly useful where there is a large amount of
serole
routine.
Conveniences for making the determination by method, such as 2-minute sand glass for regulating time of boiling, test plate, dropping bottles
Soxhlet's
for ferrocyanide solution
pj g
and
acetic acid, are
shown
in
166 tubes for
Filter
deter-
Fig. 167. Violette's
Method.
The volumetric method
of
mining su s ars
-
reducing
copper reduction, which is used most extensively in France, is that of Violette. The proportions of copper sulphate, Rochelle salts and alkali employed in the Soxhlet method may be used in the Violette
determination, or the Violette solution
*
may
be taken which consists of
Wiley's "Agricultural Analysis" (1897), III, 130.
t Ibid.
Journal of Analytical Chemistry, 4 (1890), p. 427.

Sidersky's
"Manuel"
(1909), p. 95.
394
SUGAR ANALYSIS
Fig. 167.
Ross's
method
for determining reducing sugars.
36.46 gms.
CuS0 4 .5
H 0,
2
200 gms. Rochelle
salts
and 500 gms. sodium
hydroxide solution of 1.2 sp. gr. made up to 1000 c.c. The Violette solution takes a slightly larger amount of copper sulphate than the Soxhlet solution in order that 1 c.c. may correspond to
5 = 5.263 mgs. the invert sugar derived from 5 mgs. of sucrose or || of invert sugar. The ratio of invert sugar and copper Sulphate for the Soxhlet and Violette solutions is accordingly 5 34.64 :: 5.263 36.46.
in
preferred by some chemists for convenience sucrose the method of inversion and copper reducdetermining by
Violette solution
is
The
tion.
The end point
of the reduction in Violette's
method
is
determined,
as in the early process of Barreswil, by the disappearance of blue color from the copper solution. The details of the method are as follows:
Ten
cubic centimeters of the mixed copper solution are transferred
to a large test tube 20 to 22 mm. in diameter and 22 to 24 cm. long; 5 c.c. of distilled water are added in case the solution is rich in reducing
sugars and a few small pieces of pumice stone, which have been ignited and then washed in acid and water. The copper solution is then heated to boiling, the grains of pumice stone giving a smooth ebullition
and preventing the sudden ejection
of liquid
from the tube.
The sugar
395
solution to be tested, which should have been previously clarified and diluted to about 0.5 to 1.0 per cent invert sugar, is then added from a
burette, a few cubic centimeters at a time, the copper solution being As the reduction proceeds boiled for 2 minutes after each addition.
the blue color of the solution becomes more of a reddish violet, due to the diminishing intensity of the blue and the increasing amount of the
Towards the end of the reduction it is necessary red cuprous oxide. to hold the tube against a white wall or paper and observe the color of
the clear solution, after the red oxide begins to settle. When the final drop of sugar solution discharges the last trace of blue color, the reading of the burette is noted, and the calculation of sugar made as previously described.
A little practice is required in the Violette method in following the disappearance of the blue color. The chemist should standardize his solution against invert sugar, following the same procedure in determining end point as in making an analysis. The Violette method is much simpler than the Soxhlet method and is for this reason preferred by many chemists. The Soxhlet method, on
the other hand, owing to the more sensitive method of determining the end point of reduction, has a much greater degree of accuracy. The Violette method has been modified by Spencer,* so as to include the ferrocyanide test for unreduced copper. Some chemists have improve the method by employing larger test tubes and The possibilities of modiusing 20 c.c. of the mixed copper solution.
also sought to
fication in this direction are of course unlimited
and do not require
special description.
Another volumetric process, using the disapPavy's Method. pearance of blue color as end point, is the method of Pavy,f which is based upon the fact that when Fehling's solution is reduced in presence of ammonia the precipitated cuprous oxide is dissolved as a colorless
any unreduced copper being indicated by the characteristic cuprammonium compounds. The disturbing influence of the precipitated cuprous oxide upon the color of the solution is thus avoided and, in the absence of air, the change from blue to colorless at the end point becomes quite sharp.
solution,
blue color of the
Pavy's
copper
2
solution
is
prepared
as
follows:
34.65
gms.
0, 170 gms. Rochelle salts and 170 gms. potassium hydroxide are dissolved in water to 1000 c.c. It is preferable, however,
as in Soxhlet's
* t
CuS0 4 .5H
method
to
make up
for
the copper and alkali-tartrate solu(1906), p. 131.
Spencer's
"Handbook
Cane Sugar Manufacturers"
Pavy's "Physiology of the Carbohydrates" (London, 1894), p. 71.
396
tions separately to
SUGAR ANALYSIS
500
c.c.,
and to mix equal quantities of the two just mixed copper solution are transferred to a liter flask, 300 c.c. of ammonia of specific gravity 0.880 are added and the volume completed to 1000 c.c. 20 c.c. of the ammoniacal Fehling's
before using; 120
c.c.
of the
solution are reduced
by
The reduction
is
0.01 gm. glucose. carried out in a flask of about 150 c.c. capacity,
provided withâ two-hole stopper, one opening of which is connected with the tip of the burette containing the sugar solution and the other with a bent glass tube for the escape of air and steam (Fig. 168).
Fig. 168.
Pavy's method for determining reducing sugars.
in the flask,
Forty cubic centimeters of the ammoniacal copper solution are placed and after inserting the stopper the solution is brought to a boil. The sugar solution is then added at the rate of 60 to 100 gentle
drops per minute, the discharge being regulated by a
Pavy pinch cock

(C); the ebullition
397
must be maintained without
interruption.
is
When
the blue color begins to lighten, the sugar solution drop until the last trace of color is just discharged.
added drop by The end point is
made more
plate (P).
sensitive
by looking through the
solution against a white
The reduction must be made
in complete absence of air, otherwise
the dissolved cuprous oxide will be reoxidized. used to prevent the entrance of air, through
A precaution sometimes
momentary
cooling,
is
to
use a bent-glass exit tube, fitted with a rubber valve, dipping into a beaker of water. Care must also be taken not to prolong the time of
reduction, otherwise all the oxide not be dissolved.
ammonia
will
be expelled and the cuprous
In Pavy's method 1 molecule of glucose reduces 6 molecules of cupric oxide instead of 5 molecules as by Fehling's solution. These proportions vary somewhat, however, according to concentration and
The solution should, therefore, be other conditions of experiment. standardized against glucose or invert sugar following the exact method
employed
in analysis.
Pavy's method gives good results, when the reduction is carried out with complete exclusion of the air. The extra precautions necessary
for
making the determination, and the
failure of the
method
to give
good results with colored solutions, have prevented the process from becoming generally employed. Conversion Tables for Volumetric Determination of Sugars. The calculation of reducing sugars by any of the volumetric methods
is
much
simplified
by the use
of appropriate conversion tables.
If
volume of Fehling's solution be taken, which always corresponds to a fixed amount of reducing sugar, as, for example, 0.5 gm. in Table LXX, and the sugar solution for titration be made up so as to contain this same amount of substance (as 0.5 gm.) in 1 c.c., then the formula for determining reducing sugar becomes
_
:
0.5 0.5
100
XV
100
in
which
is
the per cent of reducing sugar in the substance and
V the
cubic centimeters of sugar solution necessary for the reduction. If the substance be very high or very low in reducing sugar, an even fraction or multiple of 0.5 gm. may be taken for the amount of sub-
stance to be dissolved in
1 c.c.
Thus
for 0.05
gm. of substance
in 1 c.c.
R =
-' and
for 1
gm. of substance in
1 c.c.
R =
398
SUGAR ANALYSIS
of analysis a table giving different
Under the above conditions
multiples of the reciprocals of the burette readings will give the corThe following example responding percentages of reducing sugars.
will illustrate
the method for constructing such a table.

Fehling's solution taken
0.2
gram
of reducing sugar
Volume
of
sugar solution
for reduction.

within this interval until the exact
the reducing sugar
is
399
amount necessary
for oxidizing all
found.
for this
The
pipettes
employed
1 c.c.
lower part from
to 5 c.c.
graduated into hundredths.

to 0.01 c.c.
method are graduated in their and in the stem contain an extra 1 c.c. With three trials and employment of the
ferrocyanide test, the volume of Fehling's solution can be determined The following example illustrates the application of the
method.
First trial.
400
SUGAR ANALYSIS
Soxhlet* Variability in Reducing Power of Monosaccharides. showed that when a solution of glucose acted upon Fehling's solution the first portion added reduced most strongly and the succeeding porThis variability in reducing power is found to tions gradually less so.
be different, however, for the monosaccharides, glucose, fructose, invert sugar, galactose, etc., than for the disaccharides, lactose and maltose. As examples of the variability in reducing power of monosaccharides the following results are given. The values, which were calculated
from Bertrand's sugar tables, represent the milligrams of copper duced by each succeeding 10-milligram portion of added sugar.
TABLE LXXI
Shouting variability in reducing power of monosaccharides
re-
Number
of series.

k are easily determined empirically, and knowing these construct tables for any of the reducing sugars.
it is
401
possible to
As an example of this method of calculation the following values are taken from the experimental work of Allihn *
:
No.
of series
(ra)
25
10 mgs. of glucose reduce 18.0 mgs. copper 20 mgs. of glucose reduce 38.2 mgs. copper 250 mgs. of glucose reduce 463.0 mgs. copper.
18.0
38.2
18.0
20.2
= c. = d.
. .
.
Substituting the above values for c and d in the equation for n = 25, - (25 - 2 25 - 3 = 463.0 18 (25 1) 20.2 ) k
whence k
0.14.
The equation 18
(n
1)
20.2
-(n-2 + n-3+n
n) 0.14 will
give the milligrams of copper reduced by any multiple under the conditions of Allihn's experiments.
of 10 mgs. of glucose
Suppose
of glucose.
it is
required to find the milligrams of copper reduced by 100 mgs.
- 1) 20.2 (10 2 -10 - 3 = 194.8 mgs. Cu. 18 (10 ) 0.14 Allihn obtained by actual experiment 195 mgs. of copper by the reducing action of 100 mgs. of glucose.
Calculation of Reduction Tables.
The
calculation of tables for
the copper-reducing power of different sugars is usually made method of least squares, according to the general formula:
by the
y
in
=A
+ Bx + Cx\
which x is the milligrams of copper reduced by y milligrams of sugar and A, B and C constants. Having determined by experiment the values of x for 10 or more values of y, the calculation of A, B and C is made in the same manner as described on page 175.
As an example
quoted.
of the method of least squares the work of Allihn is again Allihn found that different amounts of glucose under constant con-
ditions of experiment reduced the following
amounts
of copper.
Mgs. Mgs.
of glucose (y) of copper (x)
402
Variability in
SUGAR ANALYSIS
The variability Reducing Power of Disaccharides. lactose is of maltose and different from that noted for power the monosaccharides. According to the amount of free alkali, time of boiling and other conditions, succeeding portions of maltose and lactose, while usually showing a slight loss, may show either no change at all, or even a slight gain in reducing power over preceding portions of the same This peculiarity of maltose and lactose is explained by a slight sugar.
in reducing
hydrolysis of the sugar into monosaccharides of higher reducing power. A slight inversion of this kind takes place with sucrose, which is strictly speaking a non-reducing sugar, and it no doubt occurs to a greater or
less
extent with
all
higher saccharides
upon
boiling with Fehling's solu-
tion.
illustration of the reducing power of successive portions of the maltose, following results are taken from the tables of Wein and of
As an
Munson and Walker.

TABLE LXXII
Showing
variability in reducing
power of maltose
Number
of Series.

Methods and
Allihn;
of copper reduced
403
tables for estimating different sugars from the amount from Fehling's solution have been devised by Soxhlet;
Wein; Meissl; Herzfeld; Lehmann; Kjeldahl; Pfluger; Ost; Honig and Jesser; Brown, Morris and Millar; Bertrand; Defren; Munson and Walker; Kendall; and many others. It is impossible to describe all these processes and only a few of the more typical methods will be selected. The method of Allihn,* which is one of the widest illustrates well the various principles involved and will be known, described first in somewhat fuller detail. Allihn's Method for the Determination of Glucose. The followof details Allihn's method with the of several ing description processes for determining the amount of reduced copper are taken from the Methods of Analysis of the Association of Official Agricultural
Chemists, f
PREPARATION OF REAGENTS
Copper-sulphate Solution.
Dissolve 34.639 gms. of
CuS04.5H2
in
water and dilute to 500
c.c.
Alkaline-tartrate Solution. Dissolve 173 gms. of Rochelle salts and 125 gms. of potassium hydroxide in water and dilute to 500 c.c.
DESCRIPTION OF METHOD
the copper solution, 30 c.c. of the alkaline-tartrate solution and 60 c.c. of water in a beaker and heat to boiling. Add
c.c. of
Place 30
25
c.c. of
prepared as not to contain
the solution of the material to be examined, which must be so more than 0.250 gm. of glucose, and boil for
Filter immediately exactly two minutes keeping the beaker covered. through asbestos without diluting, and obtain the weight of copper by one of the methods described in the following section. The correspond-
ing weight of glucose
is
found from Allihn's table (Appendix, Table
10).
METHODS FOR DETERMINING REDUCED COPPER

"Filter the Reduction of the Cuprous Oxide in Hydrogen.| cuprous oxide immediately through a weighed filtering tube made of hard glass, using suction. Support the asbestos film in the filtering tube
with a perforated disk or cone of platinum, and wash free from loose Provide the fibers before weighing; moisten previous to the filtration. tube with a detachable funnel during filtration, so that none of the
precipitate accumulates near the top,
* J.
where
it
could be removed by
prakt.
Chem.
[2],
22, 46.
S.
t Bull.
t Ibid.
107 (revised), U.
Bur. of Chem., pp. 49-53.
404
SUGAR ANALYSIS
ii
in
Fig. 169.
Forms
of tubes for filtering cuprous oxide.
Fig. 170.
Showing methods of
filtering
cuprous oxide with
filter
tube or
Gooch
crucible.
405
the cork used during the reduction of the cuprous oxide. Transfer all the precipitate to the filter and thoroughly wash with hot water, following the water by alcohol and ether successively. After being dried, connect the tube with an apparatus for supplying a continuous current of dry hydrogen, gently heat until the cuprous oxide is completely re-
duced to the metallic
state, cool in the current of
hydrogen and weigh."
Several forms of tubes for filtering cuprous oxide are shown in Glass wool is sometimes used in place of a platinum disk for Fig. 169.
holding the asbestos, but makes a less resistant support (see Fig. 169 III).
Fig. 171.
Apparatus for reducing cuprous oxide to copper. A, hydrogen generator; B and C, gas driers; D, filter tube containing cuprous oxide.
A
is
shown in and all the
convenient method of filtering cuprous oxide by means of suction A continuous filtration should be maintained Fig. 170. precipitate should be transferred to the tube before the
liquid
above the asbestos is allowed to run completely through. Too rapid or too irregular filtration may cause particles of cuprous oxide to run through the asbestos. A fine jet of water will usually bring all the cuprous oxide into the filter tube; should any of the precipitate remain
adhering to the beaker a feather, or a rubber-tipped rod, will assist the removal. The reduction of the cuprous oxide to copper by means of hydrogen All air must be expelled from the tube before is shown in Fig. 171.
406
heating, otherwise there
SUGAR ANALYSIS
is is
continued until
all
water
danger of explosion. The heating should be A desiccator of the expelled from the tube.
is
form shown in Fig. 172

ing
filter
convenient for holdfilter
The
tubes before weighing. asbestos used for loading the

is
tubes
should be of a kind which hot Fehling's solution. of preparation used by

is
not attacked by
The following method Munson and Walker *
recommended.
Preparation of Asbestos. Prepare the asbestos which should be the am phi bole
variety
by
first
digesting with
hydrofree
chloric acid for
two or three days.
Wash
from acid and digest for a similar period with soda solution, after which treat for a few hours with hot alkaline copper-tartrate solution of
Fig. 172.
filter
Desiccator for
tubes.
Then wash the
the strength employed in sugar determinations. asbestos free from alkali, finally
for several hours, for use.
In preparing filter tubes or Gooch crucibles load with a film of asbestos one-fourth inch thick, wash this thoroughly with water to remove fine particles of
after
washing
free
digest with nitric acid from acid shake with water
and
asbestos; finally wash with alcohol and ether, dry for 30 minutes at 100 C., cool in a desiccator and weigh. It is best to dissolve the cop-
per with nitric acid each time after weighing and use the same felts over and over again, as they improve with use.
The method of estimating copper by reduction of the precipitated cuprous oxide, although not so exact as the electrolytic method, is In the nevertheless sufficiently accurate for most purposes of analysis.
case of
impure sugar products the cuprous oxide is frequently contaminated with mineral or organic matter, and in such cases the
results.
method gives too high
Determination of Reduced Copper by Electrolysis. Deposition Filter the cuprous oxide in a Gooch from Sulphuric-acid Solution.^ crucible (as shown in Fig. 1 70) wash the beaker and precipitate thoroughly with hot water without any effort to transfer the precipitate to the filter. Wash the asbestos film and the adhering cuprous oxide into the beaker
,
by means
refilter
After the copper is all in solution, through a thin film of asbestos and wash thoroughly with hot
*
J.
of hot dilute nitric acid.
Am. Chem.
Soc., 28, 666.
t Bull.
107 (revised), U. S. Bur. of Chem., pp. 49-53.

water.
407
of sul-
Add
10
c.c. of
dilute sulphuric acid, containing
200
c.c.
phuric acid
(sp. gr.
1.84) per liter,
and evaporate the
filtrate
steam bath until the copper salt has largely crystallized. fully on a hot plate or over a piece of asbestos board until the evolution Add of white fumes shows that the excess of nitric acid is removed. from 8 to 10 drops of nitric acid (sp. gr. 1.42) and rinse into a platinum dish of from 100 to 125 c.c. capacity. Precipitate the copper by elecWash thoroughly with water, alcohol and ether successively, trolysis. If preferred the electrolysis can be dry at about 50 C. and weigh. conducted in a beaker, the copper being deposited upon a weighed
platinum cylinder. Deposition from Sulphuric- and Nitric-acid Solution.*
Filter
on the Heat care-
and
wash
Transfer the asbestos film from the as previously described. crucible to the beaker by means of a glass rod and rinse the crucible
c.c.
with about 30
of a boiling mixture of dilute sulphuric
acids, containing 65 c.c. of sulphuric acid (sp. gr. 1.84)

nitric acid (sp. gr. 1.42) per liter.
and nitric and 50 c.c. of

is
Heat and
agitate until solution
completed;
filter
and
electrolyze.
Filter and wash as preDeposition from Nitric-acid Solution.^ Transfer the asbestos film and adhering oxide to viously described. Dissolve the oxide still remaining in the crucible by the beaker.
means of 2 c.c. of nitric acid (sp. gr. 1.42), adding it with a pipette and receiving the solution in the beaker containing the asbestos film.
filter,
Rinse 'the contents of the beaker until the copper is all in solution, dilute the filtrate to a volume of 100 c.c. or more and electrolyze.
a nitrate solution
When
should be
made with water
the nitric acid
may
all
washing of the deposit acidulated with sulphuric acid in order that be removed before the current is interrupted.
is
electrolyzed, the first
Leach's Electrolytic Apparatus.
convenient apparatus for
the electrolytic deposition of copper in sugar analysis is that of Leach shown in Fig. 173. A is a hard rubber plate 50 cm. long and 25 cm.
wide provided with four insulated metal binding posts B, each carrying at the top a thumb screw by which a coiled-platinum-wire electrode may be attached. In front of each post is a copper plate about 4 cm. square covered with thin platinum foil P, which is bent around the
edges of the copper plate and so held in place, the copper plate being screwed to the rubber from beneath. On the square platinum-covered plate is set the platinum evaporating dish which holds the solution
*
Bull. 107 (revised),
U.S. Bur. of Chem., pp. 49-53.

(1911), p. 608.
t Ibid.
j Leach's
"Food Inspection and Analysis"
408
SUGAR ANALYSIS
from which the copper is to be deposited, the inside of the dish forming the cathode, while the coiled platinum wire, dipping below the surface In front of each platinum-covered of the solution, forms the anode.
plate
is
a switch S, and at either end of the hard-rubber plate
is
a bind-
ing post R, for connection with the electric current.
Fig. 173.
Leach's electrolytic apparatus for determining reduced copper.
Four determinations may be carried on simultaneously in four platinum dishes, if desired, the wiring and the switches being so arranged that beginning at one end of the plate either the first dish, or the first two or the first three, may be thrown in or out of the circuit at will without interrupting the current through the remaining dishes. A cover with wooden sides and glass top fits closely over the whole ap,
paratus as a protection from dust, but may easily be lifted off to manipulate the dishes when desired. The sides of the cover are perforated to
permit the escape of the gas formed during the electrolysis. The ordinary street current is used when available, and the strength of the current may be varied within wide limits by means of a number
of 16- or 32-candle-power lamps
K, coupled
in multiple,
and a rheostat
409
L, consisting of a vertical glass tube sealed at the bottom, containing a column of dilute acid, the resistance being changed by varying the
length of the acid column contained between the two platinum terminals immersed therein, one of which is movable. A gravity battery of four cells may be employed if the laboratory is not equipped with
electric lights.
In using the apparatus the plating process should go on till all the is deposited, which requires several hours or over night with a current of about 0.25 ampere. Before stopping the process the absence
copper
of copper in the solution should be
proved by removing a few drops ammonia, then acetic acid and testing with ferrocyanide of potassium. If no brown coloration is produced, all the copper has been plated out. Throw the dish out of circuit by means of
with a pipette, adding
first
the switch, pour out the acid solution quickly before it has a chance to dissolve any of the copper, wash the dish first with water and then with
alcohol, dry
nitric acid.
and weigh.
dish
The copper may be removed from the platinum

The
by strong
is
electrolytic process for determining reduced copper

all
the most
exact of
methods.
The determination, however,

and
is
involves a conlittle
siderable expenditure of time sugar laboratories where there
for this reason is
but
used in
a large amount of routine. Peters * has devised a rapid elecElectrolytic Method of Peters. trolytic method for the determination of copper, whereby the metal
deposited from an alkaline-tartrate solution, such as is used in The electrolysis is carried out either preparing Fehling's solution. in platinum dishes placed upon plates of sheet brass to which the
is is made, or in glass beakers or large test tubes, which case large cylindrical strips of sheet copper may be used for the cathode. The anode consists of a flat or cylindrical spiral of platinum wire, which should be placed at a distance of 1 cm. or less A volume of 10 c.c. copper solution from the cathode surface. (which may be slightly acid or alkaline) is usually taken, to which is added an approximately equal volume of a solution containing 35 gms.
cathode connection
in
pure Rochelle salts and 25 gms. potassium hydroxide (purified by For copper solutions containing free sulphuric alcohol) in 100 c.c. or nitric acid, two volumes of the alkaline-tartrate solution may be
solution
0.4 to 1.0 c.c. of a saturated aqueous potassium-cyanide then added according to the amount of copper present; the amount of cyanide solution should be less than sufficient to dis-
used.
From
is
J.
Am. Chem.
Soc., 34, 426.
410
SUGAR ANALYSIS
charge the blue color. If the copper deposit should be found to be too soft or dark colored, more cyanide should be used; an excess of the latter, however, greatly lengthens the time for complete deposition
of the copper.
In making the determination the direct 110- volt current of a lighting system is used with three 32-candle-power lamps interposed as
under these conditions the voltage measures 2.6 and the During the electrolysis the solution is warmed by a amperage small burner placed under the brass plate to one side of the cathode vessel; if test tubes are used they are placed upon wire gauze over a
resistance;
2.85.
The evolution of gas and the circulation of warm cause a liquid very rapid deposition of copper, which is usually comin less than 30 minutes. The solution should be covered during plete
small flame.
electrolysis to
To determine
prevent loss by spraying. the completion of electrolysis, Peters recommends
the Endemann-Prochazka * hydrobromic acid test. One volume of concentrated sulphuric acid is diluted with 2 to 3 volumes of distilled
About 1 c.c. of the dilute acid is placed in a narrow test tube, a few crystals of potassium bromide added and the whole heated to A drop of the solution to be tested is then added; as small boiling.
water.
an amount as 0.007 mg. copper

If
will cause a red color to develop. the deposition of copper is complete, the solution in the cathode vessel, without breaking the current, is displaced by a small stream
of water until the resistance
lamps are extinguished; under

solution.
cedure no copper
deposit of copper
is
lost
by
is
then washed
this proelectrode containing the in alcohol and ether, dried and
The
weighed. On account of the similarity in composition of the electrolyte employed by Peters to that of the alkaline-tartrate solution used in
Allihn's
method, the process recommends
itself for
the determination
of copper in the original Fehling's solution or in the filtrate
from the
reduced cuprous oxide obtained in the analysis of sugar solutions. Several volumetric processes have been devised for determining copper in the precipitate of cuprous oxide. Of these the permanganate,
the iodide and thiocyanate methods will be described.
oxide as in the previous methods.
Volumetric Permanganate Method, f Filter and wash the cuprous Transfer the asbestos film to the beaker, add about 30 c.c. of hot water and heat the precipitate and asbestos thoroughly. Rinse the crucible with 50 c.c. of a hot saturated
*
Chem. News, 42, 8. t Bull. 107 (revised), U. S. Bur. of
Chem., pp. 49-53.
411
solution of ferric sulphate in 20 per cent sulphuric acid, receiving the
After the cuprous rinsings in the beaker containing the precipitate. oxide is dissolved, wash the solution into a large Erlenmeyer flask and
nate.
immediately titrate with a standard solution of potassium permangaOne cubic centimeter of the permanganate solution should equal 0.010 gm. of copper. In order to standardize the permanganate solution, make six or more determinations with the same sugar solution,
titrating one-half of the precipitations
and determining the copper
in
the others
by
electrolysis.
electrolysis,
divided
The average weight of copper obtained by by the average number of cubic centimeters of
for the titration,
is
permanganate solution required
equal to the weight
of copper equivalent to 1 c.c. of the standard The reaction between the ferric sulphate
permanganate solution. and cuprous oxide is ex-
pressed as follows:
Fe2 (SO 4 ) 3
Since
+ Cu
+ H S0
2
FeS0 4
+ 2 CuSO + H 0.
4
2
1 atom, or 16 parts, of is required to oxidize the iron reduced 2 atoms, or 127.2 parts, of Cu, and 1 c.c. of n/10 permanganate contains 0.0008 gm. of active 0, then 1 c.c. of n/10 permanganate is
by
equivalent to 0.00636 gm. Cu. For a solution containing 5 gms. of potassium permanganate to the liter, 1 c.c. will be equivalent very
gm. of copper. Owing to slight deviations in practice from the above theoretical equation, the copper value of the permanganate must always be determined by direct experiment. Volumetric Iodide Method,* Low's Modification.^ Standardization of the Thiosulphate Solution. Prepare a solution of sodium thiosulclosely to 0.01
phate containing 19 gms. of pure crystals to 1000

rately about 0.2
c.c.
Weigh accu-
gm. of pure copper foil and place in a flask of 250 c.c. Dissolve capacity. by warming with 5 c.c. of a mixture of equal volumes of strong nitric acid and water. Dilute to 50 c.c., boil to expel the red fumes, add 5 c.c. strong bromine water and boil until the bromine is thoroughly expelled. Remove from the heat and add a
slight excess of strong
ammonium
hydroxide (about 7
is
c.c.
of 0.90 sp. gr.).
shown by a change Again Now add a slight exof color of the liquid, and a partial precipitation. cess of strong acetic acid (3 or 4 c.c. of 80 per cent acid) and boil again Cool to room temperature and for a minute to redissolve the copper. add 10 c.c. of a solution of pure potassium iodide containing 300 gms.
boil until the excess of
ammonia
expelled, as
For a
analysis see paper

f J.
study of the iodide method for determining copper by Peters, J. Am. Chem. Soc., 34, 422. Am. Chem. Soc., 24, 1082.
critical
in sugar
412
SUGAR ANALYSIS
Titrate at once with the thiosulphate of potassium iodide to 1000 c.c. solution until the brown tinge has become weak, then add sufficient
Continue the tistarch liquor to produce a marked blue coloration. tration cautiously until the color due to free iodine has entirely vanThe blue color changes toward the end to a faint lilac. If at ished.
this point the thiosulphate
be added drop by drop and a
little
time be
allowed for complete reaction after each addition there is no difficulty in determining the end point within a single drop. One cubic centi-
meter of the thiosulphate solution 0.005 gm. of copper.
will
be found to correspond to about
After washing the precipitated cuprous Determination of Copper. the Gooch crucible with a watch glass and dissolve the oxide, cover
nitric acid (1 1), poured under the Catch the filtrate in a flask of 250 c.c. capacity, wash watch glass and crucible free of copper; 50 c.c. of water will be sufficient. Boil to expel red fumes, add 5 c.c. of bromine water, boil off the bromine and proceed exactly as in standardizing the
oxide
by means
of 5 c.c. of
warm
watch glass with a pipette.
thiosulphate.
In a later modification of the above method, Low has found it possible to dispense with the use of bromine, the nitrous acid being
expelled from the copper solution by boiling, adding ing, acidifying with acetic acid and again boiling.
reaction between the copper acetate expressed as follows:
ammonia, heatis
The
and potassium iodide

2
3 2
2
Since
1
Cu(C 2 H 3 O 2 ) 2
and
1 c.c.
+ 4 KI =
Cu2 I2
+ 4 KC H
+I
2.
atom, or 63.57 parts, of copper liberates
parts, of iodine
of n/10 thiosulphate solution (24.8 gms.
atom, or 126.92 Na2 S 2 Oa
5 H2 to 1000 c.c.) reacts with 0.01269 gm. I,then 1 c.c. n/W thiosulphate corresponds to 0.00636 gm. Cu. For a solution containing 19.5 gms. of pure sodium thiosulphate to the liter, 1 c.c. will be equivalent very closely to 0.005 gm. of copper. In actual practice the above reaction does not proceed with absolutely quantitative precision, the results of the determination varying somewhat according to concen-
tration of acid, excess of reagents, temperature and other conditions. It is, therefore, important always to standardize the thiosulphate
solution against pure copper under the exact conditions which are followed in analysis.
Kendall's Modification of the Iodide Method. The removal of the nitrous acid, formed in dissolving the copper, is the chief difficulty in the iodide method. Kendall * has modified the method by removing
*
J.
Am. Chem.
Soc., 33, 1947.

the nitrous acid with hypochlorite, the free chlorine, which being afterwards removed with phenol.
is
413
evolved,
oxide, after filtering and washing upon a Gooch crucible, is dissolved in 10 to 15 c.c. of 30 per cent nitric acid into a 300 c.c. Erlenmeyer flask. The volume of solution and washings
The cuprous
should be between 50 and 60

centrated nitric acid;
5
c.c.
c.c.
with an acidity of 4 to 5
c.c.
con-
sodium hypochlorite solution are then added of such strength that the. iodine liberated by 5 c.c. is equivalent The solution is allowed to stand to 30 c.c. of n/10 thiosulphate.
of
2 minutes, when 10 c.c. of a 5 per cent colorless phenol solution are quickly added. The chlorine gas above the liquid is removed by blowing into the flask and the sides are washed down with a jet of
then made slightly alkaline with sodium with acetic acid; 10 c.c. of 30 per cent potasacidified sium iodide solution are then added and the free iodine titrated with standard sodium thiosulphate, as under Low's modification, using soluble starch as indicator. The thiosulphate is previously standardized under the same conditions as those of the method. against pure copper In working with known weights of copper between 20 and 340 nigs., Kendall found the error of his method to exceed in no case 0.3 mg.
water.
The
solution
is
hydroxide and
Peters' s Modifications of the Iodide
Method.
Peters* has found that
boiling the nitric-acid solution of copper in the presence of talcum powder will remove completely all lower oxides of nitrogen and leave
the solution, after cooling and diluting, in suitable condition for titration. The copper, or its compound, is dissolved in an Erlenmeyer
flask in the least possible
one-half
its
volume
of
volume of concentrated nitric acid, to which water has been added; 5 to 10 c.c. of dilute
less, of
acid are sufficient for 0.5 gm., or
copper.
After solution 15 to
pure powdered talcum are added, and the mixture boiled vigorously for 5 to 10 minutes. After cooling to room temperature distilled water is added and 10 c.c. of a saturated
25
c.c.
of distilled
water and a
little
potassium-iodide solution, the dilution being so regulated that the final volume of liquid at the end of the thiosulphate titration is about 120 c.c.
Peters has also employed the iodide
method
in the determination
of copper in the alkaline-tartrate solutions, or filtrates, occurring in
sugar analysis.
In the modification employed, 20
c.c.
of Allihn's
alkaline-tartrate solution, 20 c.c. of Fehling's copper-sulphate solution and 20 c.c. of water (as in a blank determination), or of the aqueous
reducing-sugar solution, were taken, making the total volume for * J. Am. Chem. Soc., 34, 422.
414
reduction always 60
filtered,
c.c.
SUGAR ANALYSIS
washed and the
After the reduction the cuprous oxide is which has a volume of 70 to 75 c.c.,
After cool-
nitrate,
acidified with 4 to 5 c.c. of concentrated sulphuric acid.
iodide are added ing to about 20 C., 10 c.c. of saturated potassium and the solution titrated with standard thiosulphate in the usual
way.
The end point of the titration in the iodide method is best mined according to Peters by noting the point at which a drop
deterof the
thiosulphate solution ceases to produce a perceptible white area upon the quiet surface of the titration liquid. As in the case of all other modifications of the iodide method, the thiosulphate solution must be standardized against pure copper under the exact conditions of
the analysis.
Potassium iodide is an expensive reagent and where many determinations of copper are made by this method, the waste titration liquids and cuprous iodide precipitates should be saved for recovery
of the iodine.
Volumetric Thiocyanate
lowing solutions are required:
Method
(Volhard-Pfluger).*
The
fol-
(a) n/W silver-nitrate solution, (6) n/10 ammonium-thiocyanate solution, (c) a cold saturated solution of sulphur dioxide (S0 2 ) in water, (d) nitric acid of sp. gr. 1.2, free from nitrous acid, (e) a saturated solution of ferric alum, (/) normal sulphuric-acid
solution.
tube, or Gooch crucible, containing the cuprous oxide is weighed and the approximate amount of copper determined. The cuprous
The
filter
oxide
then dissolved from the asbestos with nitric acid, the solution treated with a slight excess of normal sulphuric acid solution (/) necessary
is
to convert
the copper into copper sulphate and evaporated to dryness. The copper sulphate is then dissolved in water and washed into a 300-c.c. graduated flask. Sodium carbonate solution is added to the point of
all
turbidity and then 50 c.c. of the sulphurous acid reagent solution is boiled for 1 minute and then n/10 thiocyanate
until there
is
(c).
The
added
.
(6)
an excess of about 5
is
c.c.
above the calculated amount
necessary for precipitating the copper as cuprous thiocyanate
Cu2 (SCN) 2
The
then cooled, made up to 300 c.c., shaken and filtered filter Should the first runnings appear turbid, they through dry paper. are returned to the filter; 100 c.c. of the clear filtrate are diluted with 100 c.c. of water, 50 c.c. of nitric acid (d) and 10 c.c. of ferric-alum solution
solution
(e) are added, and the solution titrated with n/10 silver nitrate (a) until the red color is discharged. The addition of silver solution is continued
*
Pfliiger's
Archiv, 69, 423.

to the next
415
back
even number of
c.c.,
and then the solution
titrated
with n/10 thiocyanate until the white liquid just begins to turn pink. Let A be the cubic centimeters of n/10 thiocyanate added to the 300 c.c. of solution, B the cubic centimeters of n/10 silver nitrate added
to the 100 c.c. of nitrate,
cyanate to titrate
Since
1 c.c.
and C the cubic centimeters of n/10 back excess of B.
thio-
n/10 thiocyanate = 6.357 mgs. copper then the total milligrams of copper (Cu) are found by the formula Cu = 6.357 (A +3 C 3 B) The thiocyanate solution should be standardized against pure copper under the conditions of analysis, as in the permanganate and iodide methods. Of other volumetric processes which Volumetric Cyanide Method. reduced are used for determining copper may be mentioned the wellThe unreduced known cyanide method. copper in the filtrate from the
.
cuprous oxide is titrated with standard potassium cyanide solution until the blue color disappears. The difference between the copper in the volume of Fehling's solution taken, and that found in the filtrate after
reduction,
is the amount of copper reduced by the sugar. In this Determination of Copper by Weighing as Cupric Oxide. method the cuprous oxide, after collecting upon a Gooch crucible, is heated to redness for about 15 minutes, when it is converted to black To insure complete oxidation care must be taken that cupric oxide.
the oxide
not exposed to the reducing action of the illuminating gas during ignition. For this reason the operation is best carried out in a
is
muffle.
If porcelain Gooch crucibles are used they should have open bottoms with loose perforated disks for supporting the asbestos (CaldwelPs
crucible, Fig. 174).
The
one-piece porcelain
Gooch
crucible
ignition.
is
liable to crack at
high temperatures of
is
Finely-divided cupric oxide
hygroscopic and,
after cooling in a desiccator, should be
weighed as
The weight of cupric oxide Fig. 174. Gooch cruquickly as possible. cible with detachthe factor 0.7989 gives the weight of multiplied by
metallic copper.
Several sugar tables, as KjeldahPs

results in
and Defren's, express

used.
the labor of calculation,
when
this
terms of cupric oxide, thus avoiding method of determining copper is
is
The method of estimating copper from the weight of cupric oxide one of the most accurate of the indirect methods. With impure prohowever, the precipitate of cuprous oxide frequently carries
ducts,
416
SUGAR ANALYSIS
down mineral matter and this contamination will impair somewhat the accuracy of the method (see Table LXXIII).
Determination of Copper by Direct Weighing of the Cuprous In this method the precipitated cuprous oxide is collected Wash thoroughly in a filter tube or Gooch crucible in the usual way. with hot water, then with 10 c.c. of alcohol and finally with 10 c.c. of ether. Dry the precipitate 30 minutes in a water oven at tlie temOxide.
perature of boiling water; cool and weigh. The weight of cuprous oxide multiplied by 0.8882 gives the weight of metallic copper. The
sugar tables of Munson and Walker express results in terms of cuprous oxide, and the use of these tables will save much labor of calculation
when
this method of determining copper is used. Direct weighing of the cuprous Contamination of Cuprous Oxide. oxide is the simplest of the gravimetric methods for estimating reduced copper in sugar analysis. The process, however, is less accurate than
the other methods previously described.

sults
The method
gives good re-
with sugar solutions of high purity, but with impure products the cuprous oxide is contaminated with mineral and organic impurities, which may affect considerably the accuracy of the determination.
extent of the error in estimating copper from the weight of cuprous oxide is shown by the following comparative analyses made by Sherwood and Wiley * upon a variety of sugar-containing products.
The
TABLE LXXIII
Comparison of Methods for Determining Reduced Copper

The
results
417
of
upon the molasses residuum show a contamination
the cuprous oxide with organic matter as shown by the differences in copper as calculated from the suboxide and oxide, and with mineral
matter as shown by the differences in copper as calculated from the oxide and by the volumetric method.
solutions of pure sugar and such liquids as beer, where the matter consisted largely of carbohydrates, the calculation of organic from the weight of cuprous oxide gave accurate results. In the copper case of the malt extracts, which contained added peptones, the precipitated cuprous oxide seemed to carry down a considerable amount of albuminoid matter from solution; in the case of the molasses the precipitated copper seemed to be in partial combination with certain nitrogenous bases such as xanthine.
With
Similar comparisons
upon methods
of determining copper in the
analysis of cane-sugar products are given in
Table
LXXX.
usually able to form an opinion of the purity of the oxide from its physical appearance. If the precipitate is yelcuprous low or greenish-red in color, or has a flaky appearance, there is evidence
is
The chemist
which case the reduced copper must be determined of the one direct methods. by
of contamination, in
CAUSES AFFECTING THE ACCURACY OF ESTIMATING SUGARS FROM A DETERMINATION OF REDUCED COPPER
In addition to the errors in determining reduced copper, there are a of other causes which affect the accuracy of the analytical
this class.
number
methods belonging to
Purity of
A frequent cause of inaccuracy in deterthe methods of copper reduction is the presence of mining sugars by mineral or in the Fehling's solution. The copper organic impurities the caustic alkali and sulphate, especially the Rochelle salts should be
Reagents.
of the purest quality.
The copper sulphate and
alkali-tartrate solu-
tions should be filtered separately through glass wool, or asbestos, and the mixed reagent should be perfectly clear and show no trace of cuprous
oxide after boiling.
blank determination should be made upon each
fresh lot of solution; the crucibles, or filter tubes, used in the blank test should show no increase in weight under the conditions of experi-
ment followed
Degree
in analysis. of Dilution and
Time
of Boiling.
The
effect of
varying
the dilution of Fehling's solution, or the time of boiling, is shown by the following comparison of results from Allihn's table with those
418
SUGAR ANALYSIS
Koch and Ruhsam's
modifications of
obtained by Wein's, and by

Allihn's
method.

The
this
419
results
show
for
pure invert sugar a slight tendency towards
increase in reducing
power with increase in pressure; the error due to cause, however, is slight and may be neglected for ordinary at-
mospheric conditions. When sucrose is present the increase in pressure causes a marked increase in the amount of reduced copper, owing to the
much
greater degree of inversion.
Surface Area of Solution.

the Fehling's solution
is
reduced copper. solution to the air,

of
The diameter of the vessel in which heated has been found to influence the amount With wide beakers, which expose a larger area of
more cuprous oxide is lost by oxidation (through being redissolved in the alkaline-tartrate solution) than in narrower beakers. Kjeldahl has eliminated the error due to oxidation by making the reduction in an atmosphere of hydrogen or of oxygen-free illumi-
nating gas. Under the same set of conditions the oxidation error
is
a constant
one and the discrepancies due to this cause are eliminated by making the reduction always in beakers of the same size. A 350-400-c.c. lipped beaker of Jena, or Non-sol glass, 7-8 cm. in diameter is about
the proper
size.
SPECIAL COPPER-REDUCTION METHODS

Modifications of Allihn's Method.
Allihn's method gives the solutions upon sugar containing 0.4 to 1.0 per 0.10 to 0.25 gm. glucose in the 25 c.c. of solution.
is
most accurate
cent glucose,
results
i.e.,
than 50 mgs. of glucose are present the method show wide discrepancies in the hands of different chemists.
less
When
apt to
Several
modifications of Allihn's method, involving a longer period of heating, have been devised for the purpose of increasing the accuracy of the
determination with dilute sugar solutions. Pfliiger's Method. Pfliiger,* who uses the same reagents and volumes of solutions as in Allihn's method, has modified the determination
by heating the mixed sugar and
Fehling's solutions (145
c.c. in all)
bath for exactly 30 minutes and then diluting with 130 c.c. of cold water before filtering. The cuprous oxide is filtered upon asbestos and, after washing and drying, the weight of precipitate determined. Owing to the frequent occurrence of impurities in the cuprous oxide, especially when working with fluids or extracts of animal origin,
in a boiling- water
Pfliiger advises to
make
*
also a direct determination of the copper
by
means
of the thiocyanate method.

Pfliiger's
Archiv, 69, 399.
420
Pfliiger's table giving
SUGAR ANALYSIS
the weights of glucose corresponding to difand copper, is found in the Appendix
ferent weights of cuprous oxide
(Table 11).
Koch and Ruhsam's
Method.
In this modification the same
reagents and volumes of solutions are used as in Allihn's and Pfliiger's methods. The mixed sugar and Fehling's solutions (145 c.c. in all)
are first brought to a boil and then set in a boiling-water bath for exThe solution without diluting is then filtered through actly 30 minutes. asbestos in a Gooch crucible and the reduced copper determined by any
of the usual
methods.
The
glucose table for
Koch and Ruhsam's method
is
given in the
Appendix (Table 12). Koch and Ruhsam's modification was designed for determining glucose in tannin extracts, etc., and is the official method of the American Leather Chemists and other similar associations.
The modifications of Allihn's method, using 30-minute heating, are considerably more accurate than the original process upon dilute glucose solutions and should be employed for determining small amounts
of sugar in urine, tannin extracts and other animal and vegetable substances of low glucose content. When, however, the 25 c.c. of sugar solution contain over 0.10 gm. of glucose, Allihn's original method of
2-minute boiling
siderable
be followed with perfect safety, and with a conThe fact that more copper is reduced upon longer heating does not affejt the accuracy of the method, since the tables were standardized under exactly similar conditions.
may
economy
of time.
Application of Allihn's Allihn's ducing Sugars.

the
Method to the Determination of Other Remethod has been employed for determining
other reducing sugars besides glucose. Honig and Jesser f have used method for determining fructose and have constructed a table givthe ing copper-reducing power of fructose for different weights of sugar.
In Table
the fructose values of Honig and Jesser, and the corresponding glucose values of Allihn, are given for several weights of reduced copper. The ratio of fructose to glucose, for the same weight
of copper, is also given.
LXXIV
is
For equal weights of sugar the amount of copper reduced by fructose about 92 per cent of that reduced by glucose. Soxhlet found by his
(p.
volumetric method
391) that for equal weights of sugar the reducing

of glucose.
power of fructose was 92.4 per cent that

*
J. Soc.
Chem.
Ind., 13, 1227.
t Monatshefte, 9, 562.

TABLE
421
LXXIV
Showing Comparative Reducing Power of Fructose and Glucose

Reduced copper.
422
of glucose
SUGAR ANALYSIS
reduced 2.205 gms.
CuO and
=
61.
gm.
of maltose 1.345 gms.
CuO.
The
relative copper, or cupric oxide, reducing power of maltose
would then be
K=
100
In the examination of starch-conversion products the copper-reducing power of maltose, expressed by the symbol R, is sometimes 'Sullivan the taken as the standard. Taking the previous values of
of glucose
would be =
100
164.
*
In place of the constant K, Brown, Morris and Millar

stituted the value
K,
have subrela-
which
is
K.
According to this system the

is
tive copper-reducing
K.
power
of maltose (using O'Sullivan's results)

for the
0.61
of
The
values for
K,
when determined
same absolute weights
the two sugars, are practically identical with the reducing ratios as calculated in the previous section.
reduce 100 mgs.
Thus from Defren's table for glucose and maltose 44.4 mgs. of glucose CuO and 44.4 mgs. of maltose reduce 61.1 mgs. CuO then
=
-JlUU
0.611, K for maltose.
Using again Defren's table 44.4 mgs. glucose and 72.8 mgs. maltose reduce 44 4 respectively 100 mgs. CuO, then ^ = 0.610, the reducing ratio of maltose to
7 .G
glucose.
however, is calculated from the weights of sugars as determined solution factor 3.86, as is sometimes done, then the true reducing ratio is not found unless a correction be applied as indicated on page 32.
If K,
by the
The
same constancy
disaccharides, lactose and maltose, do not show usually the in reducing ratios for different weights of copper as the
monosaccharides.
This is due to the partial hydrolysis of the disaccharides as previously explained; the reducing ratio is usually higher the greater the amount of disaccharide. The copper-reducing ratios of
lactose
and maltose are approximately as follows
for Allihn's
method:
=
LactoJhydrate
66 to
71 or approximately 0.7.
'
~
jyF~TT*
=
J.
0-56 to 0.62, or approximately 0.6.

Soc. (1897), 96,
Chem.

If
423
the copper-reducing power of a sugar is determined (as by Allihn's method), the corresponding glucose value of Allihn's table divided by the reducing ratio of the sugar to glucose will give the weight of sugar
in the
25
c.c.
of solution.
c.c. of
25 Example. mgs. of copper.
a fructose solution gave by Allihn's method 265.3
The amount of glucose corresponding to 265.3 mgs. of copper, according to Allihn's table, is 137.45 mgs. 137.45 -f- 0.915 (the reducing ratio of fructose The amount of fructose corresponding to to glucose) = 150.2 mgs. of fructose.
265.3 mgs. of copper according to
Honig and Jesser
is
150 mgs.
The reducing ratios of the different sugars, have an important bearing upon the analysis of sugar mixtures, as described in Chapter XVI. Special copper-reduction methods and tables, similar to those of
for other reducing sugars. It is imin all of these detail and only the following examples describe possible to are given for invert sugar, maltose and lactose. The methods and
Allihn,
have been established
tables are taken
from Wein's "Zuckertabellen."
for Determining Invert Sugar. The Soxhlet formula for Fehling's solution is used; 25 c.c. of the copper-sulphate solution and 25 c.c. of the alkaline-tart rate solution are mixed with the sugar solution, which should not contain over 0.245 gm. of invert
Meissl's *
Method
sugar.
is added to make the whole up to 100 c.c., the heated to boiling and kept at ebullition for exactly 2 minutes. liquid The cuprous oxide is then filtered on asbestos and the reduced copper
Enough water
is
determined by any of the usual methods. The amounts of invert sugar corresponding to different weights of reduced copper are given in the Appendix in Table 13, which was calculated by Wein from Meissl's
reduction factors.
Wein'sf Method for Determining Maltose.

for Fehling's solution
is
The Soxhlet formula
and
0.25
of the copper-sulphate solution 25 c.c. of the alkaline-tartrate solution are mixed and heated to
c.c.
used; 25
boiling;
which should not contain over and the liquid boiled for exactly added gm. 4 minutes. The cuprous oxide is filtered on asbestos and the reduced The amounts of copper determined by any of the usual methods.
25
c.c.
of the sugar solution,
of maltose, are then
maltose corresponding to different weights of reduced copper are given

in the
Appendix
*
in
Table
14.
According to
Brown, Morris and Millar, J whose results have been
Z. Ver. Deut. Zuckerind., 29, 1050. Wein's "Tabellen." % J. Chem. Soc., Trans., 71, 96.
424
SUGAR ANALYSIS
confirmed by Ling and Baker,* the table of Wein gives results which are about 5 per cent too low. The Soxhlet formula Soxhlet'sj Method for Determining Lactose.
for Fehling's solution
and 25
c.c.
of
is used; 25 c.c. of the copper-sulphate solution the alkaline-tartrate solution are mixed with 20 to
100 c.c. (according to concentration) of the milk-sugar solution, which should not contain over 0.300 gms. of lactose hydrate. If less than 100 c.c. of milk-sugar solution is taken sufficient water is added to make
the whole up to 150
asbestos
c.c.
The
liquid
is
at ebullition for exactly 6 minutes.
of the usual and the The amounts of lactose hydrate corresponding to different weights of reduced copper are given in the Appendix in Table 15, calculated by Wein from Soxhlet's reduction factors.
The cuprous oxide reduced copper determined by any
then heated to boiling and kept is filtered on
methods.
UNIFIED COPPER-REDUCTION METHODS FOR SEVERAL SUGARS
The confusing
fact that different investigators
multiplicity of copper-reducing tables is due to the have confined their work to one single
sugar for one individual set of conditions. A number of chemists, however, have worked with the purpose of establishing one uniform method for all reducing sugars. Examples of such unified methods are those of
Kjeldahl and Woy, Defren, Munson and Walker, and Bertrand. Unified Method of KjeldahlJ and Woy. In Kjeldahl's method, as modified by Woy, the Fehling's solution is prepared for each analysis
with a freshly weighed portion of Rochelle

tions are used:
salts.
The
following solu-
are dissolved to 1000 c.c. 2 (A) 69.278 gms. of pure CuS0 4 .5 (B) 130 gms. of pure sodium hydroxide (the amount must established by titration) are dissolved to 1000 c.c.
be
c.c.
According to the richness of the sugar solution, 15

of
c.c.,
30
c.c.
or 50
mixed reagent are made up

c.c. c.c.
in
an Erlenmeyer
A,
flask holding
about
150
For 15 For 30
of reagent take 7.5 c.c. of

salts.
c.c.
7.5 c.c. of
B B
and 2.6 gms.

and
5.2 gms.
Rochelle
c.c. of reagent take 15.0 Rochelle salts.
of
A, 15.0
c.c. of
For 50
c.c. of
reagent take 25.0

salts.
c.c. of
A, 25.0
c.c.
of
and 8.65 gms.
Rochelle
The sugar
* J.
solution
is
then added, the total volume of liquid in the

I
Chem.
Soc., Trans., 71, 509.
t J. prakt.
Chem.,
21, 266.
Neue Z. Riibenzuckerind., 37, 29. Chem. Centralblatt. 97 [2], 986.

flask being
425
always brought to 100 c.c. The flask is then plunged in a boiling-water bath and heated for exactly 20 minutes, while leading
through the liquid a stream of hydrogen, or of illuminating gas which has been freed of oxygen by passing through a gas washer containing The reoxidation of the pyrogallic acid and sodium hydroxide solution. the air is in this oxide by way prevented. At the end of the 20 cuprous minutes the cuprous oxide is filtered on asbestos, washed, ignited and
weighed as cupric oxide. The amounts of glucose, fructose, invert sugar, lactose hydrate or maltose corresponding to different weights of cupric oxide or copper are given in the Appendix in Table 16, which was calculated by Woy for the 15-c.c., 30-c.c. and 50-c.c. volumes of reagent. The Kjeldahl-Woy method is one of great exactness, being carried out under rigidly defined conditions. The rather complicated details
in preparing the copper reagent
and
in conducting the reduction
have
prevented the process from coming into extensive use. Unified Method of Brown, Morris and Millar.* In this method, which is adapted from a previous process by O'Sullivan, the Fehling's
solution
is prepared by dissolving 34.6 gms. crystallized copper sulphate, 173 gms. Rochelle salts and 65 gms. anhydrous sodium hydroxide to 1000 c.c.; 50 c.c. of the reagent are placed in a beaker of about 250 c.c.
capacity and of 7.5 cm. diameter. The beaker is set in a boiling-water bath, and when the solution has acquired the same temperature, the measured volume of sugar solution is added and the whole made up
The beaker is covered with a to 100 c.c. with boiling distilled water. The clock glass, returned to the bath and heated exactly 12 minutes.
cuprous oxide
is filtered
in a
tube and weighed as metallic copper or
Brown, Morris and Millar (Appendix, Table 17) gives the weight of copper and cupric oxide which correspond to the same weight of glucose, fructose and invert sugar, the order of arrangement being the reverse of that in most tables.
In Defren's method, which Unified Method of Defren.f adapted from O'Sullivan, Soxhlet's formula for Fehling's solution
used; 15
c.c. of
is
cupric oxide. The table of
is
the copper-sulphate solution and 15 c.c. of the alkalinetartrate solution are diluted with 50 c.c. of water in a 300-c.c. Erlenmeyer flask. The latter is then immersed for 5 minutes in a boiling-
water bath, when 25

burette.
minutes.
run in from a heated for The flask is replaced in the bath and exactly 15 The cuprous oxide is then filtered on asbestos, washed,
c.c.
of the sugar solution are quickly
J.
t J.
Chem. Soc., Trans., 71, 281. Am. Chem. Soc., 18, 751.
426
SUGAR ANALYSIS
The amounts of glucose, maltose ignited and weighed as cupric oxide. or lactose corresponding to different weights of cupric oxide are given in the Appendix in Table 18.
Unified
Method
and
of
Munson and Walker.*
Transfer 25
c.c.
each
alkaline-tartrate solutions (Soxhlet's formula) to a 400-c.c. Jena or Non-sol beaker and add 50 c.c. of reducing sugar- soluof the copper
tion, or,
if
make the final volume 100
a smaller volume of sugar solution be used add water to c.c. Heat the beaker upon an asbestos gauze
over a Bunsen burner; so regulate the flame that boiling begins in 4 minutes, and continue the boiling for exactly 2 minutes. Keep the beaker covered with a watch glass throughout the entire time of heating.
Without diluting
filter
the cuprous oxide at once on an asbestos
felt in
a porcelain Gooch crucible, using suction. Wash the cuprous oxide thoroughly with water at a temperature of about 60 C., then with 10 c.c. of alcohol and finally with 10 c.c. of ether. Dry for 30 minutes
in a
oxide.
water oven at 100 C., cool in a desiccator and weigh as cuprous The amounts of glucose, invert sugar, lactose or maltose cor-
responding to different weights of cuprous oxide or copper are given in the Appendix in Table 19. Unified Method of Bertrand.f The following formula is used in preparing the copper reagents:
(A) 40 gms. of pure CuS0 4 .5 2 are dissolved to 1000 c.c., (B) 200 gms. of Rochelle salts and 150 gms. of sodium hydroxide in sticks are dissolved to 1000 c.c.:
H0
20 c.c. of the sugar solution, which should not contain over 0.100 of reducing sugars, are transferred to a 150-c.c. Erlenmeyer flask,
gm. and
and added. The liquid is then heated to The soluboiling and kept at gentle ebullition for exactly 3 minutes. tion is then filtered through asbestos, the precipitate of cuprous oxide
c.c.
20
each of solutions
washed with
distilled water and the reduced copper determined by the volumetric permanganate method. The table of Bertrand (Appendix, Table 20) gives the different
weights of reduced copper which correspond to the same weight of invert sugar, glucose, galactose, maltose and lactose, the order of
arrangement being the same as in the table of Brown, Morris and

Millar.
METHODS FOR DETERMINING REDUCING SUGARS IN PRESENCE OF SUCROSE

Reference has been
Fehling's solution
*
J.
made
upon the higher saccharides.
to the slight hydrolytic action of hot While this action in

f Bull, soc. chim., 35, 1285.
Am. Chem.
Soc., 28, 663; 29, 541; 34, 202.

case of sucrose
is
427
slight it
is,
nevertheless, sufficiently pronounced to
cause a considerable error in the determination of reducing sugars when much sucrose is present.
ing's
Conditions Affecting the Reducing Action of Sucrose upon FehlThe reducing action of sucrose upon Fehling's Solution.
solution is proportional first, to the concentration of the sucrose and, If enough reducing second, to the amount of copper left unreduced. sugars are present to precipitate nearly all the copper from the FehlThis is ing's solution the inversion of the sucrose will be very slight.
shown
in Table LXXV, which gives a series of experiments by Browne.* Constant quantities of sucrose, and varying amounts of glucose, were taken, and a determination of the latter made by Allihn's method.
TABLE
Showing Influence of Sucrose
A,
Sucrose taken
in 25 c.c.
LXXV
the
Upon
Reducing Action of Glucose
428
SUGAR ANALYSIS
Table
ing for the reducing action of sucrose, when using Allihn's method. gives a comparison of the actual errors and of the results
LXXV
corrected
by means
of such a formula.
In the volumetric methods of Soxhlet, Violette, etc., where the invert sugar solution is added to the point of complete reduction, no
excess of copper
is left
in solution,
and the
error
due to the presence
of
sucrose
is
practically negligible.
of special copper-reduction
number
for determining invert sugar in sugar-house products.
methods have been designed The methods
are classified according to the excess of sucrose over invert sugar in the material to be analyzed. Herzf eld's* Method for Determining Invert Sugar in Raw Sugars
Containing Less than 1.5 per designed for the analysis of the solution, which should contain free from suspended or soluble
follows
:
cent Invert Sugar.
This method
c.c.
is
higher grades of raw sugar. 20 gms. of material in 100

impurities,
is
The sugar
and be
conveniently prepared as
graduated
Dissolve 44 gms. of sugar in about 100 c.c. of water in a 200-c.c. flask. A little normal lead-acetate solution, just sufficient
is
for clarification,
then added and the volume completed to 200
c.c.
shaken, filtered and 100 c.c. of the filtrate (22 gms. Sufficient carbonate, or sugar) measured into a 100-110 c.c. flask. sulphate of sodium is then added to precipitate the excess of lead and the volume made up to 110 c.c. The solution is shaken, filtered and
solution
is
The
50
c.c.
Heat 25
of the filtrate (10 gms. of sugar) used for the determination. c.c. each of the copper-sulphate and alkaline-tartrate solu-
tions (Soxhlet 's formula) to boiling; the 50 c.c. of clarified sugar solution are then added and the whole boiled for exactly 2 minutes. The cuprous
oxide
is
filtered
by any
on asbestos, washed and the reduced copper determined of the usual methods. The amounts of invert sugar corre-
sponding to different weights of copper are given in the Appendix, in Table 21. In case the percentage of invert sugar in the raw sugar exceeds
Herzf eld's method is no longer applicable. Meissl and Wein'st Method for Determining Invert Sugar in Mixtures of 90 to 99 per cent Sucrose with 10 to i per cent Invert
1.5 per cent,
This method is designed for the analysis of low-grade raw or of other sugar-house products which do not contain a sugars, large
Sugar.
*
Z. Ver. Deut. Zuckerind. (1885), 985. Wein's "Tabellen."
429
excess of invert sugar. The sugar solution is prepared as in the previous final the filtrate method, being diluted if necessary so as not to contain
more than
Mix
25
0.2 to 0.245 gms. of invert sugar in 50 c.c. c.c. each of the copper-sulphate and alkaline- tartrate solu-
tions (Soxhlet's formula) with the 50 c.c. of clarified sugar solution; the liquid is then heated to boiling and kept at gentle ebullition for exactly
2 minutes.
The cuprous oxide
is
then
filtered
on asbestos, washed and
the reduced copper determined by any of the usual methods. For determining the weights of invert sugar corresponding to different weights of reduced copper, for percentages of sucrose between 90
and
99, the following
condensed table has been calculated by Wein.
Intermediary values can be easily calculated
by
interpolating.
TABLE
LXXVI
(Meissl and Wein.)
For Determining Invert Sugar in Presence of Sucrose.
(S)
In mixtures of sucrose and invert sugar (/) in parts per hundred.
430
SUGAR ANALYSIS
304.7
then for the intermediary 324.0 mgs. of copper

324.0
357.7
15Q Q
304.7
mgg
of invert sugar or 1.59 per cent.
method is not applicable to products which conmore than 10 parts invert sugar in 100 parts of mixed sugars. For this reason the method has largely given place to the more general process of Meissl and Hiller. Meissl and Killer's * Method for Determining Invert Sugar in Mixtures Containing less than 99 per cent Sucrose and more than i
Meissl and Wein's
tain
per cent Invert Sugar.

all
is designed for the analysis of The the sugar-house products except highest grades of raw sugars. method is based upon the principle of taking such a quantity of material
This method
for analysis that the invert sugar will reduce nearly all the copper,
thus reducing the error due to presence of sucrose to a minimum. The sugar solution is prepared as in the two previous methods so
and deleading, contain 20 gms. of in large test tubes by adding a series of solutions sample. Prepare 4 and 5 c.c. of each tube successively. Add this solution to 1, 2, 3,
that 100
c.c.,
after clarification
c.c.
of the
to boiling 2 minutes,
mixed copper reagent (Soxhlet's formula) to each, heat and filter. Note the volume of sugar solution
which gives the filtrate lightest in tint, but still distinctly blue. Place 20 times this volume of the sugar solution in a 100-c.c. flask, dilute to the mark and mix well. Use 50 c.c. of the solution for the determinaThe tion, which is conducted as in the method of Meissl and Wein. invert sugar is then calculated -by means of the following formulae. Let Cu = the weight of copper obtained;
P=
W = the weight of
F=
-rt-
the polarization of the sample; sample in the 50
c.c.
of solution
used for
determination; the factor obtained from the table for conversion of cop-
per to invert sugar;
= = =
approximate weight of invert sugar
=A
=
100
AX-yy
100 P
approximate per cent of invert sugar
y,
o, approximate per cent of sucrose in mixture of sugars;
100 S = /, approximate per cent of C\iF ~w~ = per cent of invert sugar.
*
invert sugar;
Z. Ver. Deut. Zuckerind. (1889), 735.
REDUCTION METHODS .FOR DETERMINING SUGARS

The
factor
431
for calculating copper to invert sugar is
then found
from the following table:

TABLE LXXVII
Meissl and Hitter s Factors for Calculating Copper to Invert Sugar for Different Ratio* of Sucrose to Invert Sugar
Ratio of
sucrose to invert sugar
7
=5:7.
432
SUGAR ANALYSIS
Munson and Walker's * Method for Determining Invert Sugar in Munson and Walker have included in their Presence of Sucrose.
unified
method
for reducing sugars determinations of invert sugar in
presence of variable amounts of sucrose. Their table (Appendix, Table 19) gives the weight of invert sugar for different weights of cuprous oxide or copper, when the total weight of invert sugar and sucrose in the
0.4 gm. amount is used 1 and 9 parts of between preferably for sugar products containing the 2.0 sucrose to 1 part of invert sugar and gms. amount for sugar
solution taken
is
0.4
gm. and 2.0 gms.
The
products containing over 9 parts of sucrose to 1 part of invert sugar. This range is sufficient to include all the products of the sugar factory. The method requires a preliminary investigation of the material in
order to determine the approximate percentages of sucrose and invert sugar for use in making up the solution.
MISCELLANEOUS COPPER-REDUCTION METHODS
amount of free alkali in Fehling's copper solution has most objectionable feature, owing to the influence which it proved has in rendering sucrose and other substances slightly copper reducing. Attempts have accordingly been made to devise a copper reagent for sugar analysis which would contain no caustic alkali. While none of the solutions thus designed has shown the same all around suitability as that of Fehling, a few of them have found a certain usefulness in
large
its
The
special cases.
Barfoed's
Copper-acetate
Method.
Barfoed's copper-acetate
solution (p. 336), which is not reduced by the disaccharides, maltose and lactose, has appealed to chemists as a convenient means of determining
glucose, fructose
ducing sugars.
and other monosaccharides in presence of the higher reBut notwithstanding its value for qualitative purposes,
attempts to use Barfoed's reagent for the quantitative determination of glucose and other monosaccharides have always given unsatisfactory
results.
Soldaini's J Copper-bicarbonate Method. Soldaini's copper-bicarbonate solution (p. 337) has also appealed to chemists as a means of avoiding certain errors resulting from tiie use of Fehling's solution. Soldaini's method, however, has usually given unreliable results, when used for quantitative purposes, the principal objections being the deposition of copper hydroxide
*
and the precipitation of lime and other mineral impurities with the reduced copper.
J.
Am. Chem.
t Z. analyt.
Chem.,
Soc., 28, 663. 12, 27.
$ Ber., 9, 1126.
433
Ost's * Copper-bicarbonate Method. Ost has modified Soldaini's reagent in order to eliminate its objectionable features. In his latest
improvement
of the method the copper reagent is prepared as follows: 250 gms. of chemically-pure potassium carbonate and 100 gms. of chemically-pure potassium bicarbonate are dissolved in water, and a solution containing 17.5 gms. of chemically-pure crystallized copper sulphate slowly added. The volume is then made up to 1000 c.c. and the solu-
tion filtered through asbestos, the first runnings of the filtrate being
rejected.
In making the determination 100 c.c. of the copper reagent are c.c. of the sugar solution and the liquid boiled for 10 minutes. The precipitate is then filtered upon asbestos and the retreated with 50
duced copper determined by any of the usual methods. Ost has unified his method for a number of reducing sugars; a few of the values for different weights of reduced copper are given in Table
LXXVIII.
TABLE LXXVIII
Showing Reducing Power of Different Sugars upon
Reduced
copper.
Ost's
Copper Solution
434
This
difficulty,
SUGAR ANALYSIS
ammonium
according to Ost, may be obviated by precipitating the oxalate during the clarification. The method
lime with
upon the whole has not offered sufficient advantages over Fehling's solution to come into general use. * Bang's Copper-bicarbonate Method. Bang has recently employed the copper-bicarbonate method for the volumetric determinaIn this method the excess of tion of very small amounts of glucose. which remains in solution after copper, reduction, is titrated with a
standard hydroxylamine-sulphate solution in presence of potassium
thiocyanate.
TABLE
Hydroxylamine.
LXXIX
435
In making the determination 10 c.c. of the sugar solution, which should not contain over 60 mgs. of glucose, are measured into a 200-c.c. flask and 50 c.c. of solution A added. The liquid is heated to boiling
and kept at ebullition for exactly 3 minutes. The solution in then cooled and solution B added from a burette until the blue color just disTable LXXIX gives the milligrams of glucose correspondappears.
ing to the cubic centimeters of hydroxylamine solution used.
Kendall's Alkaline-salicylate
Method.
Kendall* has recently
devised a method for determining reducing sugars, in which salicylic acid and potassium bicarbonate are used in place of the ordinary
alkaline-tartrate
The advantages mixture of Fehling's -solution. claimed are that the alkaline-salicylate mixture has no copper-reducing power of its own and that much larger amounts of copper are reduced by a given weight of sugar when the carbonates of the alkalies
are used in place of the hydroxides. solution is measured into a 200-c.c. Erlenmeyer flask, and the volume made up to 100 c.c. with distilled water. There are
The sugar
then added in succession 5 gins, salicylic acid, 15

solution, containing 133.33 gms.
c.c.
CuS04.5
HO
2
per
liter,
copper-sulphate and 25 c.c.
2 C03 per liter. potassium-carbonate solution, containing 600 gms. The flask is shaken until the salicylic acid has completely dissolved, and then placed in a boiling- water bath for exactly 20 minutes; the reduced cuprous oxide is then filtered upon asbestos, washed with hot water, and the copper determined by Kendall's modified iodide method (p. 412). From the milligrams of copper thus found the
corresponding weights of glucose, invert sugar, lactose hydrate and maltose hydrate are determined from a specially calculated table.
VOLUMETRIC-REDUCTION METHODS BY MEANS OF MERCURY SOLUTIONS

Of other metallic salt solutions besides copper only those of mercury have been used to any great extent for determining reducing sugars. The solution used Knapp'sf Alkaline Mercuric-cyanide Method.
in
Knapp's method is prepared by dissolving 10 gms. of pure mercuric cyanide and 100 c.c. of sodium-hydroxide solution of 1.145 sp. gr. to 1000 c.c. The solution contains 7.9363 gms. of metallic mercury per liter.
In making the determination a measured volume of the reagent, previously standardized against a known weight of the pure sugar, is heated to boiling and the sugar solution added from a burette until
a drop of the filtered solution shows upon acidifying with acetic acid no coloration with ammonium-sulphide solution. The calculation of
J,
Am, Chem.
Soc., 34, 317.
t Z. analyt.
Chem.,
9,
395.
436
SUGAR ANALYSIS
sugar is made in the same manner as described under Soxhlet's volumetric method with Fehling's solution.
The end reaction in Knapp's method has been found uncertain and the process at present is but little used. The solution of Sachsse's * Alkaline Mercuric-iodide Method.
Sachsse
is
prepared as follows:
18 gms. of pure dry mercuric iodide
(prepared by precipitating mercuric-chloride solution with potassium iodide, and washing and drying at 100 C.) are dissolved in a solution
containing 25 gms. of pure potassium iodide; a solution containing 80 gms. of potassium hydroxide is then added and the volume completed
to 1000 c.c.
The
solution contains 7.9323 gms. of metallic
mercury
per
liter.
An alkaline stannous-chloride solution, prepared by treating a solution of stannous chloride with an excess of potassium hydroxide, is
used for determining the end point. In making the determination a measured volume of reagent is heated to boiling, and the sugar solution added until a drop of the filtered solution shows no coloration with the alkaline tin solution.
The comparative reducing power

tion
is
of several sugars
upon Sachsse's
solu-
given in
Table
LXXXII,
page 474.
ESTIMATION OF HIGHER SACCHARIDES BY DETERMINING THE COPPERREDUCING POWER AFTER HYDROLYSIS
The methods previously described in this chapter for determining reducing sugars are equally applicable to the analysis of the higher nonreducing saccharides provided the latter first undergo a quantitative
hydrolysis into sugars of known reducing power. The best examples of such applications of the
Fehling's solution.
method
are the deof
terminations of sucrose, starch, dextrin and glycogen
by means
DETERMINATION OF SUCROSE BY MEANS OF FEHLING'S SOLUTION

Sucrose upon treatment with invertase or acids is hydrolyzed quan95 parts of sucrose yielding 100 parts of invert sugar. the copper-reducing power of an inverted-sucrose solution be deterwill
titatively,
If
mined, the equivalent of invert sugar multiplied by the factor 0.95

give the
amount
of sucrose present.
In making the determination care must be taken that the amount

of sugar after inversion does not exceed the limit of the tables, which for 50 c.c. of mixed Fehling's solution is about 240 mgs. of invert sugar,
*
437
or the equivalent of about 225 mgs. of sucrose. The chemist should check the method with pure sucrose, in which case the following pro-
cedure
may
be followed.
Dissolve 1.9 gms. of pure sucrose in about 75 c.c. of water in a 500-c.c. graduated flask and invert the solution according to the method
of Herzfeld, or any of the processes described in Chapter X. After cooling, the solution is nearly neutralized with sodium hydroxide (carefully avoiding any excess) and the volume completed to 500 c.c.;
of this solution (containing 200 mgs. invert sugar = 190 mgs. sucrose) are then treated according to any of the copper-reduction
50
c.c.
methods
for invert sugar
and the weight
of reduced copper determined.
The milligrams
of invert sugar, corresponding to this weight of copper, multiplied by the factor 0.95 gives the milligrams of sucrose. In applying the method to the determination of sucrose in sugar-
house products, and other substances, which contain invert sugar, the difference between the invert-sugar equivalents before and after inversion is multiplied by 0.95. The same methods for determining invert sugar should be employed in both cases. The method of calculation is best illustrated by an example:
of the
Four grams of apple must were made up to 100 c.c. (solution A). Four gms. same must were inverted, nearly neutralized and made up to 100 c.c.
(solution B).
50 50
c.c. of sol.
c.c. of sol.
104
B gave by MeissFs method 407 mgs. Cu = 230 mgs. invert sugar A gave by Meissl's method 235 mgs. Cu = 126 mgs. invert sugar Difference = 172 mgs. Cu 104 mgs. invert sugar. mgs. invert sugar X 0.95 = 98.8 mgs. or 4.94 per cent sucrose.
The mistake is sometimes made of taking the difference between the weights of reduced copper before and after inversion and calculating the invert sugar and sucrose from this. The extent of this error, which is due to the variation in the copper-reducing power for different parts of the table (as shown in Table LXXI), may be seen from the
previous example, where a difference of 172 mgs. of copper was found. 172 mgs. of copper according to Meissl's table correspond to 90.8 mgs. of invert sugar. 90.8 X 0.95 = 86.2 mgs. or 4.31 per cent of sucrose,
a result considerably less than that obtained by the other method. In calculating sucrose by any of the chemical methods, the reducing sugars before inversion must always be expressed as invert sugar, al-
though it may actually exist as glucose, lactose, maltose, etc., or a mixture of several of these. This, of course, applies only to the sucrose calculation and not to that of the reducing sugars.
438
Example.
to 500
c.c.
SUGAR ANALYSIS
(solution A).
5 gms. of a sirup containing sucrose and maltose were made up 5 gms. of the same sirup were dissolved, inverted,
nearly neutralized and
made up
to 500
c.c.
(solution B).
Maltose
-
Mgs.
Mgs.
Mga.
50
c.c. of sol.
c.c.
50
of sol.
B A
gave by Munson and Walker's method 390 gave by Munson and Walker's method 199
Difference
= =
215.0
103.7
111.3.
175.5
191
21.14 per cent sucrose in sirup. 35.10 per cent maltose in sirup. Calculating the sucrose from the difference in copper, as is sometimes wrongly done, would give the following: 191 mgs. Cu = 99.3 mgs. invert
111.3
0.95
105.7 mgs. 175.5 mgs.
= =
sugar (by Munson and Walker's table), 99. cent sucrose in sirup.
3X
0.95
94.3 mgs.
18.86 per
The unified methods and tables are most convenient for converting the equivalents of any reducing sugar into that of invert sugar. The same result, however, may be accomplished by means of the copperreducing ratios given on page 421.
Example.
10 gms. of a sirup containing sucrose and fructose were made 10 gms. of the same sirup were dissolved, inverted,
up
25 25
to 500
c.c.
(solution A).
nearly neutralized and

c.c. of sol. c.c. of sol.
made up to 500 c.c. (solution B). gave by Allihn's method 414 mgs. Cu = 221 mgs. glucose gave by Allihn's method 195 mgs. Cu = 100 mgs. glucose
Difference
=121
is
mgs. glucose.
The reducing
121
-r-
ratio of invert sugar to glucose
0.958 for Allihn's method.
0.958
126.3 mgs. invert sugar.
126.3
X
is
0.95
120 mgs.
24.00 per
cent sucrose in sirup.
The reducing
100
-^
ratio of fructose to glucose
0.915 for Allihn's method.
0.915
109.3 mgs.
21.86 per cent fructose in sirup.
Owing
sugars, as maltose
to the slight variation in the reducing ratios of some of the and lactose, it is more accurate to determine the
equivalents
by one
of the unified
methods.
DETERMINATION OF STARCH BY MEANS OF FEHLING's SOLUTION

Starch upon heating with dilute hydrochloric acid is hydrolyzed most quantitatively according to the equation (C 6 Hi 05) n -f- nH 2
al-
nC Hi2
6
6,
in
which 90 parts
of starch yield 100 parts of glucose.
The
conversion of starch into glucose may be accomplished either by direct acid hydrolysis, as in Sachsse's method, or by first converting the starch
into soluble products, as with diastase, solution with acid.
and then hydrolyzing the
filtered

Method
of
439
Sachsse, as modified by the Association of Official Chemists.* Stir a convenient quantity of the sample Agricultural from 2.5 to 3 gms. of the dry material) in a beaker with (representing water for an hour. Transfer to a filter and wash with c.c. of cold 50
250 c.c. of cold water. Heat the insoluble residue for two and a half hours with 200 c.c. of water and 20 c.c. of hydrochloric acid (sp. gr. 1.125) in a flask provided with a reflux condenser. Cool, and nearly
sodium hydroxide; complete the volume to 250 c.c., and determine the glucose in an aliquot of the filtrate by any of the usual methods of copper reduction. The weight of glucose multineutralize with
filter
plied
by 0.90 gives the weight of starch. Owing to the fact that a perfect theoretical yield of glucose is never obtained from starch by acid hydrolysis, Ost f recommends the use of the factor 0.925 for converting glucose into starch by Sachsse's
method.
method is one of the simplest processes for estimating has the objection of converting pentosans and other hemibut starch,
Sachsse's
celluloses into reducing sugars.
The method
for this reason gives too
high results in the analysis of starchy substances

cellular tissue.
which contain much
In order to eliminate this error the starch must be
tion of starch
dissolved from cellular substances before hydrolyzing with acid; solumay be effected by heating under pressure or by the
action of diastase.
Method of Determining Starch by Solution under Pressure.! Three grams of the finely-ground sample are extracted with cold water, as in the previous method in order to remove sugars, dextrin, gums, etc.
If
much
oil
or fat
is
ether.
of
The
residue
is
present the material should first be extracted with then heated in a covered flask or metal beaker,
about 200-c.c. capacity, with 100 c.c. of water in an autoclave, a form of which designed by Soxhlet is shown in Fig. 175. The heating If an autoclave is continued for 3 to 4 hours at 3 atmospheres pressure.
is
not available, Lintner pressure bottles (Fig. 176) may be used; the bottles are immersed in a glycerol bath and heated for. 8 hours at 108 to 109 C.
the digestion is finished the pressure is first allowed to subthe autoclave, or pressure flask, is opened and the solution filtered through asbestos. The insoluble residue is well washed with
side,
When
when
hot water, and should show no blue reaction with iodine

*
f
when
ex-
Bull.
107 (revised), U.
Ztg., 19, 1501.
S.

p. 221.
Chem.
Konig's "Untersuchung" (1898),
440
SUGAR ANALYSIS
The
filtrate is
amined under the microscope.

then heated with 20
c.c.
made up
to 200 c.c.
and
of hydrochloric acid, of 1.125 sp. gr., for 3
Fig. 175.
Soxhlet's autoclave.
Fig. 176.
Lintner's pressure bottle.
hours in a boiling-water bath, the flask, which holds the solution, being connected with a reflux condenser. The solution, after cooling, is nearly neutralized with
sodium hydroxide and made up to 500
c.c.
The
copper-reducing power of the solution is then determined; the glucose equivalent of the copper multiplied by 0.9 gives the corresponding equivalent of starch. Method of Determining Starch by Solution with Diastase. -
Marcker* found that the best method of dissolving starch from hemicelluloses was by means of diastase. The method of Marcker, as modified by the Association of Official Agricultural Chemists, is as follows: Preparation of Malt Extract. Digest 10 gms. of fresh, finelyground malt 2 or 3 hours at ordinary temperature wi-th 200 c.c. of
*
"Handbuch
der Spiritusfabrikation" (1886), 94.

water and
y
441
filter.
of the filtrate
tion,
Determine the amount of glucose in a given quantity after boiling with acid, etc., as in the starch determina-
in the subsequent determination. Extract a convenient quantity of the substance (ground to an impalpable powder and representing from 4 to 5 gms. of the dry material) on a hardened filter with 5 successive portions of 10 c.c. of ether; wash with 150 c.c. of 10 per cent alcohol and then with a little strong alcohol. Place the residue in a beaker with 50 c.c.
and make the proper correction
Determination.
of water, immerse the beaker in a boiling-water bath and stir constantly for 15 minutes or until all the starch is gelatinized; cool to 55 C., add
c.c. of malt extract and maintain at this temperature for an hour. Heat again to boiling for a few minutes, cool to 55 C., add 20 c.c. of malt extract and maintain at this temperature for 1 hour or until a microscopic examination of the residue with iodine shows no starch. Cool and make up directly to 250 c.c.; filter. Place 200 c.c. of the
20
filtrate in a flask with 20 c.c. of hydrochloric acid (sp. gr. 1.125); connect with a reflux condenser and heat in a boiling-water bath for two and one-half hours. Cool, nearly neutralize with sodium hydroxide
and make up to 500 c.c. Mix the solution well, pour through a dry filter and determine the glucose in an aliquot of the filtrate by any of the usual methods of copper reduction. The weight of glucose multiplied
by 0.90 gives the weight of starch. Wein * has calculated a table for the above methods which
gives
the milligrams of starch or dextrin corresponding to milligrams of reduced copper as obtained by Allihn's method. The table was constructed
table
by simply multiplying the milligrams by the factor 0.9.
of glucose in Allihn's
secures the
Of the various processes for determining starch the diastase method most perfect solution of starch with the least solution of accompanying hemicelluloses. In cases, however, where much cellular matter is present the hot water and malt solution may dissolve a small
amount of pentosans, which, by being afterwards hydrolyzed into reducing pentose sugars, introduce a slight error in the determination. A more serious error than the above consists in the incomplete hydrolysis of starch into glucose. Experiments by W. A. Noyes,f
and
his coworkers, testing the action of 2.5 per cent hydrochloric acid
show a hydrolysis, into glucose A diminished yield of theoretical. the about 97 per cent of factor somewhat greater a conversion of glucose necessitates the use
upon the malt conversion
which
is
of starch,
than
0.9.
*
Wein's "Tabellen."
t J-
Am. Chem.
Soc., 26, 266.
442
SUGAR ANALYSIS
Modification of Noyes* for Determining Starch by the Diastase In the modification recommended by Noyes the filtrate Method.
is treated with one-tenth its volume of hydro" After heating for 1 hour in a flask acid of sp. gr. 1.125. immersed in a boiling-water bath, making allowance for the time re-
from the malt digestion
chloric
quired for the solution to attain the temperature of the bath, the solu-
added to neutralize 90 per made up to a definite if on a and the filtered necessary, volume, dry filter, reducing power determined by Fehling's solution; 100 parts of glucose found in this manner represent 93 parts of starch in the original material."
tion
is
cooled,
enough sodium hydroxide
is
cent of the hydrochloric acid used, the solution
Noyes emphasizes the importance
himself with pure glucose the ratio between glucose particular solutions and method which he uses.
of each chemist determining for and copper for the
DETERMINATION OF DEXTRIN BY MEANS OF FEHLING's SOLUTION
The
principle of the
method
is
the same as that described for starch.

f a
In the process described by Konig
weighed amount of the dextrin
is
dissolved in cold water, made up to 1000 c.c. and filtered. Three in of 200 c.c. each of the filtrate are heated a bath boiling-water portions
with 20
hydrochloric acid of 1.125 sp. gr. for periods of 1, 2 and solutions are cooled, nearly neutralized with sodium hydroxide and made up to volume so that the solution does not contain over 1 per cent glucose. The glucose is then determined by any of the
c.c. of
3 hours.
The
usual methods, and the highest results of the three experiments taken as the correct value. The weight of glucose multiplied by the factor 0.9
gives the equivalent of dextrin. If sugars are also present, the glucose equivalent of these must be subtracted from the glucose equivalent after hydrolysis and the differ-
ence calculated to dextrin.
W.
The hydrolysis of dextrin by dilute hydrochloric acid was found by A. Noyes t and his co-workers to, be a little less than 95 per cent complete at the end of 2 hours' heating and the results seemed to indicate that the theoretical yield of glucose could not be obtained even by prolonged heating. The theoretical factor 0.9 for converting glucose to dextrin is no doubt considerably too low for the method of acid
hydrolysis.
*
t
J.
Am. Chem. Am. Chem.
Soc., 26, 266.

Soc., 26, 266.
Konig's "Untereuchung" (1898), p. 215.
j J.
443
DETERMINATION OF GLYCOGEN BY MEANS OF FEHLING's SOLUTION
The method is based upon the Pfluger's* Glycogen Method. of the into impure glycogen (C 6 Hio0 5 )n, which has glucose hydrolysis from the solution of animal substance. been precipitated previously One hundred grams of the finely divided tissue are heated with 100 c.c.
of 60 per cent potassium-hydroxide solution, in a boiling-water bath for 3 hours, the flask, which contains the solution, being shaken at frequent The cooled solution is made up to 400 c.c. and treated with intervals.
tion
After standing 24 hours the clear soluof 96 per cent alcohol. decanted through a filter, the precipitate of impure glycogen stirred with an excess of 60 per cent alcohol and again set aside. The in the numerous treatments the be of hastened glycogen may settling
800
c.c.
is
by adding a few drops of concentrated salt solution. The clear liquid The puriis again decanted and the process repeated for a third time. fication is then continued in the same way, twice with 96 per cent alcohol, once with absolute alcohol, three times with ether and once again with absolute alcohol. Any material adhering to the filter is then
main portion of precipitate, and the raw glycogen disThe solution is then neutralized with hydrochloric acid of 1.19 sp. gr., and transferred to a 500-c.c. flask; 25 c.c. of hydrochloric acid (sp. gr. 1.19) are then added and the liquid heated
removed
to the
solved in hot water.
in a boiling-water bath for 3 hours. The solution is then cooled, neumade to 500 filtered and the glucose determined in tralized, up c.c.,
the
filtrate by Pfluger's method. The amount of glucose multiplied by the factor 0.927 gives the corresponding amount of glycogen.
EXTRACTION OF SUGARS AND PREPARATION OF SOLUTIONS FOR CHEMICAL METHODS OF ANALYSIS
The methods and precautions previously given for the extraction of sugars and preparation of solutions for polarimetric examination hold also for the chemical methods of analysis.
Clarification
little
of
Solutions.
With products which contain but
insoluble matter, such as sugars, molasses, sirups, jellies, honeys, etc., the weighed amount of material is dissolved in water, clarified, if necessary, with a minimum of neutral lead-acetate solution, made up to
volume and filtered. The filtrate, after deleading by means of sodium carbonate, sodium sulphate, potassium oxalate or other means, as described on page 276, is then ready for analysis. With products of high purity, which contain but little mineral
matter or organic non-sugars, the use of lead acetate
*
may
be dispensed
Pfluger's Archiv,
114, 242.
444
with,
SUGAR ANALYSIS
and a few cubic centimeters
of
alumina cream be used for
clari-
fication.
Lead subPrecipitation of Reducing Sugars by Basic-lead Salts. of which are basic salts as or other lead, employed clarifying acetate, agents in the polarimetric determination of sucrose, should never be
used upon solutions in which reducing sugars are to be determined. The action of such compounds in causing a precipitation, or occlusion, of reducing sugars in the lead precipitate has already been mentioned.
Bryan found that basic-lead salts, in presence of magnesium sulphate and ammonium tartrate, precipitated in case of glucose from 3 per cent to 17 per cent, and in case of fructose from 8 per cent to 35 per cent, of the total amount of sugar in solution. Neutral lead acetate under the same conditions caused the precipitation of only 0.9 per cent of the total glucose and 0.0 per cent of the total fructose.. (See Table XL, p. 216.) In a series of independent experiments made by Bryan and Home f upon raw cane sugar and cane molasses the following results were
obtained.
TABLE
LXXX
Showing Influence of Clarification with Lead Subacetate upon Determination of Reducing Sugars
Clarifying agent and analyst.
445
Clarification with lead subacetate caused a loss of about 10 per cent of the total reducing sugars present. The variable results, due to method of estimating copper, show a contamination of the cuprous
oxide as explained on page 416. The higher results by Allihn's method are due to the greater inverting action of the more strongly alkaline
Fehling's solution.
PREPARATION OF SUGAR SOLUTIONS FROM PLANT SUBSTANCES

If the material to be analyzed contains much insoluble matter, as the case with plant substances containing cellular tissue, the sugars must first be extracted by means of water or alcohol. In the case of
is
grains, cattle-feeds, etc., the following provisional method is used by the Association of Official Agricultural Chemists.* Extraction of Sugars with Cold Water. Weigh into a flask or
20 gms. of the material, depending upon the amount of soluble carbohydrates present. Add 250 c.c. of ice-cold water, less the volume of water present as moisture in the material, and stir or shake for 4 hours. If enzymatic action is feared the extraction should be made at a low temperature, preferably by surrounding the extraction flask with broken ice; or extract at ordinary temperature with 40 to 50 per cent alcohol. If there is present in the material much soluble substance, correction should also be made for If necessary for clear filtration, the increase in volume due to solution. add from 5 to 10 c.c. of alumina cream, just before filtering. The volume of alumina cream to be added must be taken into account in determinAfter the extraction ing the amount of water used for the extraction. filter immediately, pouring back upon the filter the first portions of cloudy filtrate until the filtrate is clear. To free from soluble impurities add sufficient normal lead-acetate solution to 200 c.c. of the filtrate Remove to precipitate all impurities, make up to 250 c.c. and filter. the excess of lead by means of anhydrous sodium carbonate or anhybottle, suitable for stirring or shaking, 10 to
drous sodium sulphate, followed in the latter case by a small amount of anhydrous sodium carbonate, care being taken not to use an excess.
Filter again
and use the
clear filtrate for the determination of reducing
sugars.
extraction of sugars from plant substances by means of cold water is not always trustworthy owing to the action of enzymes upon The employment of hot sucrose, starch and other higher saccharides.
The
Bull. 107 (revised), U. S. Bur. of
Chem.,
p. 57.
446
water
is
SUGAR ANALYSIS
also often unreliable
on account of the solution of hemicelluand gums. Extraction of Sugars with Dilute Alcohol. Bryan, Given and * have recently made experiments upon the extraction of Straughn sugars from grains and similar products, using as solvents 50 per cent Both of these alcohol and 0.2 per cent sodium-carbonate solution. solvents inhibit the action of enzymes and were found to give concordant results upon certain classes of products. In many cases, however, the sodium-carbonate extraction gave much higher amounts of
loses, starch
a result, perhaps, of the solvent reducing sugar after inversion action of the alkali upon pentosans and other hemicelluloses. Bryan, Given and Straughn believe that extraction with 50 per cent alcohol, all
The method
points considered, is the most reliable method for general sugar work. outlined by them is as follows:
Place 12 gms. of the finely Method of Bryan, Given and Straughn. ground substance in a 300-c.c. graduated flask, adding, in case the material is acid, from 1 to 3 gms. of precipitated calcium carbonate. Add 150 c.c. of neutral alcohol of 50 per cent volume strength; mix thoroughly and boil on a hot-water bath for 1 hour, placing a small funnel in the neck of the flask to condense the vapor. Cool and make up to 300 c.c. with neutral 95 per cent alcohol. After mixing and settling transfer 200 c.c. of the clear solution to a distilling flask and distil The off the excess of alcohol, which is thus recovered for future use. liquid residue is evaporated to a volume of 20 to 30 c.c. (but not to dryness), and then washed with water into a 100-c.c. graduated flask. The solution is clarified with the necessary amount of neutral leadacetate solution, and, after standing 15 minutes, made up to 100 c.c. Pass through a folded filter, carefully saving all of the filtrate, to which add enough anhydrous sodium carbonate to precipitate the excess of
lead;
filter.
allow to stand 15 minutes and then pour through an ashl< Over 75 c.c. of filtrate should be obtained; 25 c.c. of the cl<
filtrate
(equivalent to 2 gms. of original material) are diluted with 25 c.c. water and used for the determination of reducing sugars; 50 c.c. oi the same filtrate are transferred to a 100-c.c. flask, inverted with 5 c.c. of concentrated hydrochloric acid, neutralized and made up to 100 c.c. Filter, if necessary, and take 50 c.c. (equivalent to 2 gms. of original
of
material)
for the determination of reducing sugars
after inversion.
The percentages of invert sugar and sucrose are usual way and the results multiplied by the factor
the volume of insoluble matter.
*
calculated in the
0.97 to correct for
Circular 71, U. S. Bur. of
Chem.

PREPARATION OF SUGAR SOLUTIONS FROM ANIMAL SUBSTANCES
Clarification.
447
Liquids of animal origin, such as blood, serum,
amounts and other nitrogenous substances which interfere with the determination of reducing sugars by the methods of copper reThe clarifying agent which is most used for such liquids is duction.
urine, milk, secretions, extracts, etc., frequently contain large of albuminoids
mercuric nitrate.
Treat 220 gms. of yellow oxide of merMercuric-nitrate Solution. cury with 300 to 400 c.c. of water; then add nitric acid in small portions, with warming and stirring, until the precipitate is dissolved. Dilute to
1000
until
c.c.
and
filter.
liquid to be clarified is treated with mercuric-nitrate solution no more precipitate forms; the solution is then nearly neutralized with sodium-hydroxide solution of 1.3 sp. gr., made up to volume and A measured portion of the slightly acid filtrate is then freed filtered. from excess of mercury by precipitating with hydrogen sulphide; the
is filtered, the hydrogen sulphide removed by a current of air and the reducing sugars determined by any of the usual methods. For the clarification of milk, the use of Clarification of Milk. and copper sulphate potassium hydroxide will be found more con-
The
solution
venient.
The
following
is
the
official
method
of the Association of
Agricultural Chemists.* Dilute 25 c.c. of the milk with 400
c.c.
of water
and add 10
cc. of
solution of copper sulphate of the strength given for Soxhlet's modification of Fehling's solution. Add about 7.5 c.c. of a solution' of
potassium hydroxide of such strength that one volume of it is just sufficient to completely precipitate the copper as hydroxide from one volume of the solution of copper sulphate. Instead of a solution of potassium hydroxide of this strength, 8.8 c.c. of a half-normal solution of sodium hydroxide may be used. After the addition of the alkali solution the mixture must still have an acid reaction and contain
Fill the flask to the 500-c.c. mark, mix and filter copper in solution. through a dry filter. Determine the lactose by any of the usual methods. In determining reducing sugars in substances of animal origin, the precipitate of cuprous oxide is often badly contaminated with mineral and organic impurities, so that the reduced copper should be determined directly and not by weighing as suboxide or oxide.
*
Chem.,
p. 119.
448
SUGAR ANALYSIS
CONCENTRATION OF SUGAR SOLUTIONS
In working with very dilute solutions, such as contain only a few hundredths of a per cent of sugar, it is often necessary to concentrate the liquid to one-half, one-fifth or one-tenth the original volume before a satisfactory determination of the copper-reducing power can be
made.
It is exceedingly important in evaporating such solutions that the liquid be kept exactly neutral, otherwise changes may result in the composition of the sugars. Traces of free acid may become sufficiently
concentrated towards the end of evaporation to hydrolyze higher saccharides,
and traces
of free alkali
be conducted in vessels which do not give up soluble alkali; the concentration of sugar solutions in glass vessels, unless of perfect resistant non-soluble quality, is for this reason to be avoided. The author has found
flasks
The evaporation
may modify or destroy reducing sugars. of solutions containing reducing sugars must
and basins
of tinned copper to be very suitable for con-
centrating sugar solutions, there being no change in reducing power after diluting and evaporating to the original volume.
If the solution to be concentrated is slightly acid an excess of finely powdered calcium carbonate (alkali free) will prevent the hydrolysis of
higher saccharides.
If
the solution,
is
alkaline, dilute acetic acid
is first
added to faint
acidity,
is
and then an excess
of calcium carbonate.
is
When
the evaporation
completed, the residue of insoluble matter
removed
by
filtration.
CHAPTER XV
SPECIAL QUANTITATIVE
METHODS
determination of sugars by means of their reducing power upon Fehling's solution, Sachsse's solution or other metallic salt combinations
is
THE
nation of particular groups of reducing sugars.
a general method, and has no value for the selective determiFor such purposes
will describe a
more
special
chapter
titative
processes of analysis must be adopted. The present number of the best known of such special quan-
methods.
DETERMINATION OF PENTOSES AND PENTOSANS Method. The methods for determining pentoses and Theory to the researches are due of Tollens,* and his school; they all pentosans
of
depend upon the conversion of the pentose sugars into furfural by
dis-
tilling with hydrochloric acid, according to the principles described on The amount of furfural, which distills over, is determined and p. 374.
calculated to pentoses. fectly to the equation,
The
yield of furfural does not correspond per-
10
CH
5
3H 0,
2
100 parts pentose
64 parts furfural
being for arabinose about 75 per cent and for xylose about 90 per cent of the theoretical. Yet by making the distillation under carefully controlled conditions, it
is possible, by means of formulae or tables which have been established for different weights of pure pentoses, to make a determination with a very close degree of approximation. Different reagents have been used for precipitating the furfural in the determination of pentoses. Tollens and Stone first attempted to determine furfural by precipitating with ammonia as furfuramide. An important advance was then made by Tollens, in company with
Giinther, de Chalmot, Flint
and Mann,
in using phenylhydrazine for
The use of phenylhydrazine was attended, precipitating the furfural. however, with certain inconveniences and was finally abandoned upon
the discovery by Councler f of the precipitating action of phloroglucin. * For a review of the subject see papers by Tollens with bibliography in Abderhalden's "Arbeitsmethoden," 1909, II, 130, and in Papier-Zeitung, 1907, Nos. 56, 60 and 61 (Reprint).
t
Chem.
Ztg., 17, 1743; 18, 966.
449
450
SUGAR ANALYSIS
The phloroglucin method, as first developed by Tollens and Kriiger,* was further improved by Tollens and Rimbach, and finally established in its present form by Tollens and Krober.f The necessary apparatus for making Description of the Method. the determination is shown in Fig. 177. From 2 to 5 gms. of substance,
according to the richness of the material in pentoses or pentosans, are placed in a 300-c.c. distillation flask with 100 c.c. of hydrochloric acid
Fig. 177.
Apparatus
for
determining pentoses and pentosans by distillation with

hydrochloric acid.
of 1.06 sp. gr. one opening of
The
which
flask
is
is
closed with a two-hole rubber stopper,
fitted to the
connecting tube* of a condenser
and the other to a small separatory funnel. The latter is preferably of cylindrical form with graduation marks at 30 c.c. and 60 c.c. The flask is then placed in a bath of Rose's alloy (1 pgh*t lead, 1 part tin and 2 parts bismuth, melting near 100 C.), which, after just
Beating
beyond the point of fusion, is brought up slightly above th6 level of the bottom of the flask. The distillate is received in a graduated
when 30 c.c. of liquid have passed over, which should require from 10 to 11 minutes, 30 c.c. more of the hydrochloric acid of 1.06 sp. gr. are added from the separatory funnel. The process is concylinder;
tinued in this
way
* t
until
fr
drop of the
distillate
shows no pink colora7.
Z. Ver.
Jour.
f.
Deut. Zuckerind., 46, 21, 195. Landwirtsch. (1900), 355, (1901),
SPECIAL QUANTITATIVE METHODS
451
tion with aniline-acetate paper (see p. 375). From 9 to 12 portions of 30 c.c. usually require to be distilled over, depending upon the amount
The distillation is then suspended and the furfural determined by precipitation with phloroglucin. Dissolve a small quantity of phloPreparation of Phloroglucin.* roglucin in a few drops of acetic anhydride, heat almost to boiling and add a. few drops of concentrated sulphuric acid. A violet color indiA phloroglucin which gives more than cates the presence of diresorcin. a faint coloration may be purified by the following method: Heat in a beaker about 300 c.c. of hydrochloric acid (sp. gr., 1.06)
of furfural.
and 11 gms.
of phloroglucin,
it
added
in small quantities at a time, stirring
constantly until
resist solution,
has almost entirely dissolved.

it is
Some
impurities
may
unnecessary to dissolve them. Pour the hot solution into a sufficient quantity of the same hydrochloric acid (cold)
but
to
make the volume 1500 c.c. Allow it to stand at least over night better several days to allow the diresorcin to crystallize out, and filter immediately before using. The solution may turn yellow, but In using it, add the volume this does not interfere with its usefulness.
containing the required
amount
to the distillate.
The distillate obtained by the Precipitation of Phloroglucide. method previously described is treated in a 500-c.c. lipped beaker with a measured volume of phloroglucin solution, so that the amount of
tion
phloroglucin is about double that of the furfural expected. The solufirst turns yellow, then green and finally becomes almost black
when
8
the amorphous dark-green precipitate of furfural phloroglucide,
begins to deposit. The liquid is then made up to 400 c.c. 12 per cent hydrochloric acid (1.06 sp. gr.) and allowed to the with over The solution, after testing with aniline-acetate stand night.
CnH O4,
paper to make sure that all furfural has been precipitated, is filtered through a weighed Gooch crucible; the precipitate of phloroglucide is brought carefully upon the asbestos and washed with 150 c.c. of water in such a way that the water is not entirely removed from the crucible is then placed upon a support, so until the y^ry last. The crucible that the bottom is free to the air, and dried for 4 hours in a boilingis then placed in a weighing bottle, cooled in a desiccator water bath;
i|
and weighed. The increase in weight is the amount of furfural phloroglucide which is calculated to furfural, pentose or pentosan according to the table of Krober (Appendix, Table 22).
of pentose in Krober's table are the averages of the corresponding weights of xylose and arabinose. The weights of pen* Bull. 107 (revised), U. S. Bur. of Chem., p. 54.
The weights
452
tosan are obtained
SUGAR ANALYSIS
by multiplying the corresponding weights
of pen-
which represents the ratio of ?iC 5 Hi O 5 to tose by of Krober has a range for weights of table The (C 5 H 8 O4) n or ijj$. 0.300 gms. For weights of phloroand 0.030 between phloroglucide Krober limits of these outside gives the formulae: glucide
the factor 0.88,
For weight of phloroglucide "a" under 0.03 gm. Furfural = (a 0.0052) X 0.5170 gm. Pentoses = (a 0.0052) X 1.0170 gm. Pentosans = (a + 0.0052) X 0.8949 gm. " " For weight of phloroglucide a over 0.300 gm.
+ +
Furfural
Pentoses
Pentosans
= = =
+ 0.0052) X 0.5180 gm. + 0.0052) X 1.0026 gm. (a + 0.0052) X 0.8824 gm.

(a (a
The factor 0.0052 represents the weight (5.2 mgs.) of phloroglucide, which remains dissolved in the 400 c.c. of acid solution. For weights of phloroglucide which exceed 0.5 gm. it may be found necessary to dry for a longer period than 4 hours in order to attain
constancy in weight. It is always better in making the determination to regulate the weight of material so that the amount of phloroglucide
falls
within the range of the table. Precautions and Limitations.
In making the determination of
pentosans by the method of acid distillation, several precautions should be noted. It is important first that the heat be applied to the flask in such a way that charring of solids upon the surface of the glass above
the liquid be avoided. Such charring is very apt to occur when the flask is heated over the open flame or upon wire gauze; the use of the
metal bath for heating is for this reason to be preferred. It is also important that the distillate be perfectly clear, and free from suspended With subimpurities, before adding the solution of phloroglucin.
much oil or wax, fatty decomposition products are sometimes carried over into the distillate; in determining pentoses in the urine of herbivorous animals, benzoic acid (a decomposition prodstances which contain
uct of hippuric acid) is distilled over in considerable amount. In all such cases the distillate must be filtered from suspended matter before
precipitating the furfural with phloroglucin. Two important limitations of the distillation
method
for determin-
1. Furfural is formed from other ing pentoses should be mentioned. substances than pentoses (the so-called furfuroids). 2. Other sub-
stances, which form a precipitate with phloroglucin, are distilled over besides furfural (the so-called furaloids).

"
Furfuroids."
453
The formation
of furfural
from glucuronic acid
The presence of glucuronic acid in urine, or of oxycellulose in plant substances, will introduce, therefore, a certain error in the determination of pentoses in
(p. 375).
and oxy cellulose has already been considered
Cross and Bevan * for this reason propose that the names furfurose, furfurosan or furfuroid be used to designate the furThe researches of Tollens show, fural-yielding complex of plants. however, that the pentosans are by far the most important of the fur-
such materials.
fural-yielding groups; the term pentosans, though not a perfectly correct expression, seems destined to remain until more accurate methods are devised for determining the different furfural-yielding groups.
The distillates obtained by boiling cellulose, starch, sucrose, fructose, glucose and other hexose carbohydrates with hydrochloric acid give with phloroglucin a small yield of phloroglucide corresponding to 0.5 to 1.0 per cent pentosans. Whether the reacting substance in such distillates
is
furfural,
been definitely determined.

into
oxymethylfurfural or mixtures of these has not A slight error is, nevertheless, introduced
the pentose, or pentosan, determination by the phloroglucin method and the chemist should always bear this fact in mind when only small amounts of phloroglucide are obtained. "Furaloids." - The distillation of other products, which give precipitates with phloroglucin, besides furfural has also been long recognized. Methylfurfural, which is obtained by the distillation of methyl-
pentoses with hydrochloric acid, forms for example a red precipitate with phloroglucin, which, unless removed by solution in alcohol, as
glucide.
afterwards described, will give too high a weight of furfural phloroIn the same way oxymethylfurfural (see p. 620) which is
slight
formed in
fructose, sucrose
amounts by the action of hydrochloric acid upon and other hexose carbohydrates, forms a precipitate
" " furaloid 7 to 23 per cent of the crude furfural. The is decomposed according to Fraps by redistilling the acid distillates; the pure furfural thus obtained is precipitated with phloroglucin, the
with phloroglucin. Frapsf has estimated that the amount of foreign products ("furaloid ") in the hydrochloric-acid distillate of different plant substances
may vary from
weight of phloroglucide corresponding to the amount of furfural-yielding bodies (pentosans or furfuroids); the difference between the
weights of phloroglucide for distillate and redistilled distillate corresponds to the amount of furaloid-yielding bodies, the exact nature of
*
Gross and Bevan's "Cellulose" (1895), p. 99.
Am. Chem.
Jour., 25, 501.
454
SUGAR ANALYSIS
Furaloid does not seem to be formed
of Barbituric Acid.
which Fraps did not determine. from the pure pentose sugars.
Precipitation of Furfural
by Means
Jager
and linger
have suggested barbituric acid
for precipitating furfural
in presence of foreign distillation products.
Cellulose, starch, sucrose
and other hexose carbohydrates give hydrochloric-acid distillates which, though reacting with phloroglucin, form no precipitate with barbituric acid. Jager and Unger claim that the reagent offers, therefore, a more accurate means of estimating pentosans.
In making the precipitation the hydrochloric-acid distillate is treated with a solution of pure barbituric acid in hydrochloric acid of 1.06 sp. gr.,
using 8 parts of barbituric acid to
solution
is
part of estimated furfural.
The
stirred
itate filtered
and after standing 24 hours the yellow granular precipinto a Gooch crucible, washed with water and dried for 4
hours at 105 C.
for the
tion.
amount
The weight of precipitate is increased by 0.0049 gm. of substance dissolved in the 400 c.c. of acid soluand barbituric acid proceeds
as fol-
The
lows:
reaction between furfural
CH
4
yCO-NH v CHO+H C ' ; CO =C H X CO-NH X

2
CH C ( ; C0+H 0. X CO-NH X
2
/CO-NHv
(206)
.
Furfural (96)
Barbituric acid (128)
Condensation product
One hundred parts

of furfural.
of condensation product thus correspond to 46.6 parts
method for determining pentosans offers several but the good features, process has not been tried sufficiently as yet by chemists to form a conclusion as to its reliability.
barbituric-acid
Jolles's
The
Method
method
from
of Determinating Pentoses.
Jolles f has recently
proposed a
particulars
for determining pentoses which differs in several that of Tollens. The substance to be distilled is
placed in a 1500 c.c. flask with 200 c.c. of 12 per cent hydrochloric acid; the flask is heated, while a current of steam is passed through the liquid, the distillation being regulated so that the volume of solution does not fall at any time below 100 c.c. By distilling the furfural with
steam the formation of humus substances
is
said to be prevented
and a
The process is continued until quantitative yield of furfural obtained. 1 c.c. of the distillate shows no coloration with Bial's orcin reagent
(p.
382);
100
*
c.c.
of the distillate (usually
between 2 and 3
liters)
Ber., 35, 4440; 36, 1222.
t Sitzungsber.
Wiener Akad., 114
(II b),
1191 (1905).
are neutralized with
to
METHODS
455
sodium hydroxide, and then made faintly acid methyl orange with a few drops of half-normal hydrochloric acid. A measured volume of TVnormal sodium-bisulphite solution is then added, and the solution allowed to stand 2 hours. The amount of bisulphite, remaining after the reaction with the furfural, is then titrated back with TVnormal iodine solution, using starch solution as indicator. The difference between the volumes of bisulphite and iodine solutions gives the amount of bisulphite which entered into combination with the furfural. The reaction between the two is expressed by the equation
:
C4H O CHO-{-NaHSO 3 = C4H 3O

3
CH /OH x
SO 3 Na
an aliquot, which is less than 5 per cent of the total a very great multiplication of any experimental involves distillate, Jolles's process has not as yet demonstrated its superiority over errors. the much shorter and simpler method of Tollens.
titration of
The
The method of Tollens for determining pentoses gives good results with pure arabinose or xylose but, as has been shown, yields only rough approximations in the case of the various furfuroids. Even in
the case of pure pentosans the calculation of furfural to a mixture of araban or xylan in equal amounts, when perhaps the pentosan itself may consist almost entirely of one substance, may involve an error of
In certain plant exudations, as several per cent in the calculation. cherry gum, the pentosans consist almost entirely of araban; in the
in the encrusting substances of
hemicelluloses of certain woods, as the beech, almost entirely of xylan; most cellular tissues of variable mix-
Until accurate methods are available for tures of araban and xylan. the estimation of xylan and araban, and for the determination of oxycellulose and other furfuroids, the calculation of furfural to a mixture
of xylan and araban in equal amounts can be regarded only as a conventional approximation.
Applications of Pentosan Method.
The determination
of pen-
tosans, notwithstanding certain limitations of the method, has found numerous applications in the assay of plant gums, in the analysis of
feeding materials, in the examination of forestry products and in other ways. A single example of such application is given in the analysis of
paper stock.
of pentosans in different
*
Krober,* for example, gives the following determinations raw materials used in paper manufacture.
Jour.
f.
Landwirtsch. (1901),
1.
456
SUGAR ANALYSIS
TABLE
Material.
LXXXI
phloroglucide
is
METHODS
457
filtered,
washed, dried and weighed in exactly the

for furfural phloroglucide.
same manner as described
The weight
either to
of
methylfurfural
phloroglucide
is
then calculated
rhamnose by the table of Ellett and Tollens or to fucose by the table of Mayer and Tollens. The rhamnose, CHaCaHgC^ H 2 O. is calculated to rhamnosan (CH3C 5 H 7 04)n by multiplying by the factor 111 = 0.80; and the fucose, CHsCsHgOs, to fucosan by the factor ||| = 0.89. The combined table giving the weights of rhamnose, rhamnosan, fucose, fucosan, and methylpentosan (mixture of equal parts rhamnosan and fucosan) corresponding to different weights
of methylfurfural phloroglucid is given in the Appendix (Table 23) Instead of the tables the following formulae may be used in which
.
Ph
is
the weight in grams of methylfurfural phloroglucide.
= Rhamnose = Methylpentosan =
Fucose
2.66
1.65
1.85
Ph Ph Ph -
12.25
1.84
6.25
Ph 2 + 0.0005. Ph 2 + 0.0100. Ph 2 + 0.0040.
Fucose decomposes slower than rhamnose with hydrochloric acid, so that the distillation must be continued longer. More decomposition in distilling fucose of are formed products methylfurfural consequently with a corresponding less yield of phloroglucide. Methylfurfural, according to Fromherz,* may also be estimated by precipitation with barbituric acid in the same manner as described for
furfural.
The
-
reaction takes place according to the equation:

2
X CH C H O CHO +H C /CO-NH CO X X
3 4
2
CO-NH
= CH C H
3
CO +H 0. CH C /CO-NH, ; , X CO-NH X
-
Methylfurfural (110)
Barbituric acid (128)
Condensation product
(220)
Two
parts of condensation product thus correspond to exactly one of part methylfurfural. The yellow crystalline precipitate is filtered in a Gooch crucible, washed with water and then dried for 5 hours in a
steam bath.
its slight
100
c.c.),
precipitate is then weighed, and after correcting for solubility in the 12 per cent hydrochloric acid (2.29 mgs. in calculated to methylfurfural by dividing by 2.
The
According to Jolles f methylfurfural may also be determined by his method of steam distillation and titration with bisulphite and iodine solutions. The reaction between bisulphite and metyhlfurfural is
similar to that described for bisulphite
and
furfural,
and the
details of
the two methods are exactly alike.

*
Z. physiol.
Chem.,
60, 241.
Ann., 361, 41.
458
SUGAR ANALYSIS
DETERMINATION OF PENTOSES AND METHYLPENTOSES IN MIXTURE
The method of determining of Tollens and Ellett. in mixture was first worked out by and methylpentoses pentoses based the and is and Tollens Ellett,* upon solubility of methylfurfural of furfural and the phloroglucide in warm 95 insolubility phloroglucide,
Method
is distilled with 12 per cent hydrochloric acid, the distillate precipitated with phloroglucin, and the mixed phloroglucides of furfural and methylfurfural filtered in a Gooch crucible, dried and weighed according to the usual process.
per cent alcohol. In making the determination the material
The crucible containing the mixed phloroglucides is then placed in a smaller beaker with 95 per cent alcohol which is heated nearly to boiling.
cible
cent
The brown-colored solution is then sucked off through the cruby means of a filter pump, and the extraction with hot 95 per alcohol repeated twice more in the same way. The crucible con-
taining the insoluble furfural phloroglucide is then dried for 2 hours in a hot-water bath and reweighed in a weighing bottle. The residual of furfural is then to calculated weight phloroglucide pentoses or pen-
tosans and the loss in weight, due to methylfurfural phloroglucide, calculated to methylpentoses, or methylpentosans, by means of the
upon known mixtures of pentoses with methylpentoses were made by Ellett and Tollens, and by Mayer and Tollens with very close agreements.
respective tables or formulae. Trials of this method of separation
who has
Modification by Hay wood of the Tollens-Ellett Method. Haywood,f recently tested the method of Tollens and Ellett, believes that
a correction should be
made
for the slight solubility of the furfural
phloroglucide in 95 per cent alcohol. Experiments made by Hay wood upon the phloroglucide obtained from pure arabinose showed that for
varying weights of substance, and extracting 3 to 5 times with alcohol, a very uniform weight of about 0.0037 gm. was always dissolved. Hay-
wood
believes the substance thus dissolved to be occluded phloroglucin
and not phloroglucide. The following slight modification of the TollensEllett method is proposed by Hay wood: Place the Gooch crucible containing the mixed phloroglucides in a 100-c.c. beaker and pour into the crucible 30 c.c. of 95 per cent alcohol
heated to 60 G. heated to 60 C.
*
Remove
Place the beaker for 10 minutes in a water bath the beaker and crucible and suck from the
Z. Ver. Deut. Zuckerind. (1905), 19. U. S. Bur. of Chem., p. 112.
t Bull. 105,

latter all alcohol
459
alternate extraction
remaining tnerein with a suction pump. Repeat this and sucking dry of the precipitate 3 to 5 times,
traction place the
After the final exaccording to the color of the nitrate obtained. Gooch crucible in a water oven and dry four hours,
making the final weighing in a closely stoppered glass weighing bottle. The difference in weight between the furfural phloroglucide plus methylfurfural phloroglucide first obtained and the furfural phloroglucide remaining after extraction with alcohol, minus 0.0037, represents the amount of methylfurfural phloroglucide present, from which
the methylpentose or methylpentosan
formulae.
is
calculated
by the
tables or
To
of methylphloroglucide
obtain the weight of pentosans, subtract the corrected weight from the weight of the mixture and calculate
according to Krober's tables or formulae.
DETERMINATION OF GALACTOSE OR GALACTAN

and his co-workers have developed a method for estimating galactose, and its higher condensation product galactan (C 6 Hi 05) n which is based upon a determination of the mucic acid formed by oxiTollens
,
dation of the substance with nitric acid.
The oxidation
of galactose to
mucic acid according to theory proceeds as follows:
C Hi
6
2HNO
H O
10
2H
+ 2NO.
Galactosg^lSO)
Mucic acid
(210)
100 partôf gatactose thus equal 116.66 parts of mucic acid. In actual experiment only about 75 per cent of the weight of galactose is obtained This yield, however, is fairly constant for the given as mucic acid. conditions of analysis, so that the weight of mucic. acid multiplied by 1J gives the weight of galactose.
The method
of Tollens as
employed by the Association of
Official
Agricultural Chemists f is as follows: Extract a convenient quantity of the substance, representing from 2.5 to 3 grams of the dry material, on a hardened filter with 5 successive portions of 10 c.c. of ether; place the extracted residue in a
beaker about 5.5 cm. in diameter and 7 cm. deep, together with 60 c.c. of nitric acid of 1.15 sp. gr., and evaporate the solution to exactly onethird its volume in a water bath at a temperature of 94 to 96 C.
After standing 24 hours, add 10 c.c. of water to the precipitate, and allow it to stand another 24 hours. The mucic acid has in the mean-
time crystallized but

*
it is
mixed with considerable material only par-
f Bull.
Ann., 227, 223; 232, 187. 107 (revised), U. S. Bur. of Chem., p. 55.
460
tially oxidized
SUGAR ANALYSIS
by the nitric acid. Filter the solution, therefore, through wash with 30 c.c. of water to remove as much of the nitric filter paper, acid as possible, and replace the filter and contents in the beaker. Add 30 c.c. of ammonium-carbonate solution, consisting of 1 part ammonium carbonate, 19 parts of water and 1 part strong ammonium hydroxide, and heat the mixture on a water bath, at 80 C., for 15 min-
The ammonium carbonate takes up the utes, with constant stirring. mucic acid, forming the soluble mucate of ammonia. Then wash the filter paper and contents several times with hot water by decant ati on, passing the washings through a filter paper, to which finally transfer the material and thoroughly wash. Evaporate the filtrate to dry ness over a water bath, avoiding unnecessary heating which causes decomposition; add 5 c.c. of nitric acid of 1.15 sp. gr., thoroughly stir the mixture and allow to stand for 30 minutes. The nitric acid decomposes the ammonium mucate, precipitating the mucic acid; collect this on a tared filter or Gooch crucible, wash with from 10 to 15 c.c. of water, then with 60 c.c. of alcohol and a number of times with ether; dry at the temperature of boiling water for 3 hours, and weigh. Multiply mucic acid by 1.33, which gives galactose and multiply this product by 0.9 which gives galactan. The method of Tollens has been used considerably by Schulze and
for determining galactan groups in different plants of the lactose Leguminosse and also by Bauer f for estimating galactose jf$ *
Steiger
in the urine.
of large amounts of foreign organic matter hinders the precipitation of mucic acid, and in case of only small amounts of the latter may prevent its separation entirely. The tendency of the
The presence
method
is,
therefore, to give too low rather than too high results.
FERMENTATION METHODS FOR DETERMINING SUGARS
A method for estimating sugars has been described (p. 299) which is based upon the change in polarization which the solution undergoes after fermenting with yeast. The fermentation methods for determining sugars are more usually carried out by weighing or measuring the carbon dioxide which is
evolved.
The theoretical yield of carbon dioxide from glucose, accordto the 2 is 48.88 per cent. 2 ing equation C 6 Hi 2 O 6 = 2 C 2 5 In actual experiments only about 45 per cent of 2 is obtained, this figure varying, however, by several per cent according to the variety
H OH + CO
CO
Landw. Vers.
Stat., 36, 11;
36, 438, 465.
t Z. physiol.
Chem.,
61, 159.
of yeast, influence of non-sugars
METHODS
461
and other conditions. The weight of carbon dioxide obtained during a normal fermentation multiplied by the factor 2.2 will give the approximate amount of fermentable hexose sugars present. The fermentation method is employed almost entirely for determining small percentages of sugar,- and has found its widest application in the determination of glucose in urine. Direct Method by Weighing Carbon Dioxide. The most accurate
method
for determining the yield of
carbon dioxide upon fermentation
Fig. 178.
Apparatus
for determining sugars

off
from weight of carbon dioxide given
by fermentation.
is shown in Fig. 178. A known amount of the solution is sterilized in a small flask, then cooled and inoculated with a pure culture of yeast. The flask is then connected by means of a condenser with a train of
absorption tubes, or bulbs. Bulb I (Fig. 178) contains a few cubic centimeters of water, the U-tubes II and III contain calcium chloride
for removing all moisture from the current of gas, the Liebig potash bulb IV, which has been previously weighed, serves to absorb the carbon dioxide, and the safety tube V, containing calcium chloride and soda lime, prevents back absorption of water, or carbon dioxide, from
the outside
air.
The fermentation
if
is
allowed to proceed either at room
temperature, or, desired, at 30 C., in which case the flask is immersed in a water bath carefully maintained at this temperature. At the end of
1 to 2 days, when no more gas passes through the bulb I, the tube V is connected with the aspirator bottle B, the pinchcock at p, which is previously closed, opened and a slow current of air, freed from carbon
dioxide
by passing through potassium hydroxide
solution, led through
462
the apparatus.
SUGAR ANALYSIS
At the end
of
an hour the liquid
in the flask is heated
nearly to boiling, while a current of cold water circulates through the condenser; in this manner the last traces of dissolved carbon dioxide
The aspiration is continued for another bulb IV is disconnected and reweighed. when the The hour, potash increase in weight gives the amount of carbonic acid. The more usual process, in the fermentation method of estimating sugars, is to estimate the carbon dioxide by measuring the volume of
are expelled from the liquid.
C. and 760-mm. atmospheric gas; 1 c.c. of evolved carbon dioxide (at to 1.96 carbon dioxide or about 4 mgs. of pressure) corresponds mgs.
glucose.
ratus
the
For determining sugars by this method special forms of appaknown as fermentation saccharometers have been devised, of which two forms devised by Einhorn and by Lohnstein are selected as
This apparatus, which
in diabetic 179.
examples. Einhorn's Fermentation Saccharometer.*

is
designed for the estimation of small
amounts of glucose urine, is shown in Fig.

of
One
commercial pressed "yeast is gram shaken thoroughly in the graduated test tube with 10 c.c. of the urine. The mixture is then poured into the bulb of the saccharometer, the apparatus being inclined so that the graduated tube is completely filled.
The saccharometer
for
is
then set aside
20
to
24
If
hours
temperature. sugar, fermentation will usually beWhen the gin in about 30 minutes.
at ordinary the urine contains
fermentation
of gas is
is
finished the
volume
measured
in the graduated
tube, the divisions of which indicate cubic centimeters of gas and also the
approximate fractions
Fig. 179.
Einhorn's fermentation
saccharometer.
glucose.
If
of per the urine contains

it
cent
than
per cent glucose
more must first
be diluted with water, the reading of the saccharometer being then For diabetic urines of straw multiplied by the degree of dilution.
color
and a
specific gravity of 1.018 to 1.022 it is
recommended
gr.
to dilute
twice; of 1.022 to 1.028 sp. gr. 5 times,

*
and 1.028 to 1.038 sp.
10 times.
Circular of information.
463
It is always desirable in making the test to make a duplicate determination upon a normal urine. The latter should show at most
only a small bubble of gas at the top of the tube; should a larger amount of carbon dioxide be obtained with normal sugar-free urine, the
yeast is probably impure and the determination should be repeated. If the suspected urine shows no more gas than the control experiment
In Lohnstein's saccharometer (Fig. 180) the liquid is fermented over mercury in a closed bulb; the carbon dioxide, which is evolved, forces
the mercury into an upright tube, the amount of displacement indicating the per cent of glucose
present.
the absence of glucose is indicated. Lohnstein's * Fermentation Saccharometer.
S
T,
is
In making a determination the detachable scale hung in position over the open end of the tube
of
and a quantity
mercury poured into the bulb

is
until its level in the tube
just opposite the
zero
mark
of the scale.
The standard weight
of
mercury, each instrument.
necessary for the adjustment,

is
accompanies
small piece of pressed yeast

its
rubbed with 2 to
;
3 times
volume
of ordinary water to a thin paste
0.5 c.c. of the urine, or other liquid to be tested, is then measured with a special pipette into the bulb;
is rinsed into the bulb with a little ordiwater and 2 to 4 drops of the yeast water nary added. The glass stopper, which should be evenly greased, is then inserted, and turned so that the small opening on its inner surface comes directly Fi g- 180 -- Lohnstein s opposite a similar opening in the stem of the bulb. fermentation sac,. ,, Any pressure of air, due to inserting the stopper, is c h a rometer thus released. The stopper is again slightly turned, so as to seal the contents of the bulb hermetically, and then securely fastened by the weight W. The apparatus is then set aside until fer-
the pipette
'
mentation is finished, which
is
indicated
by the stationary position
of the
mercury column. The length of time necessary for completing the test will depend upon the temperature but does not ordinarily exceed 1 day at 20 C.; if an incubator is available the time may be shortened con-
When fermentation is finished the siderably by fermenting at 35 C. scale division opposite the top of the mercury column indicates the
*
Miinchener med. Wochenschr.
(1899)-,
No. 50; also circular of information.
464
SUGAR ANALYSIS
percentage of sugar; for percentages of sugar below 2.0 the scale may be read to 0.01 per cent and for percentages between 2.0 and 10.0 The scale is calibrated upon one side for 20 C. and to 0.05 per cent.
upon the other

Thus:
for 35 C.; if the readings be made at intermediary the percentage of sugar is calculated by interpolating. temperatures
The reading
scale
and
3.6
of the mercury column at 25 C. was 4.0 on the 20 C. on the 35 C. scale. The corrected percentage of sugar is
then 3.6
4
'!?
oO
^
is
6
(35
ZO
25)
3.87 per cent.
Instead of finding the weight or volume of carbon dioxide the percentage of fermentable sugar may also be calculated from the amount
of alcohol
which
found by the action of yeast, or from the difference
in specific gravity of the solution before and after fermentation. valuable check upon the accuracy of the results obtained by the fer-
mentation methods
is
to determine the loss
in reducing sugars
by
means
of Fehling's solution.
COLORIMETRIC METHODS FOR DETERMINING SUGARS

of colorimetric methods have been devised for determinamounts of different sugars in solution. The first process of this kind was due to Dubrunfaut who determined small percentages of glucose by comparing the color, which was produced by heating the
A number
ing small
amounts
solution with alkalies, with the colors of solutions containing of pure glucose, which had been similarly treated.
known
In addition to the alkalies many of the special reagents, used in color and spectral reactions, such as a-naphthol, resorcin, etc., have been employed for the colorimetric estimation of sugars. The principal requirement in the use of such reagents for quantitative purposes is that the color produced must be perfectly soluble and of a The insoluble, or evanescent, colors, which fair degree of stability.
making
are produced in
many
of the reactions for sugars, are valueless for

color, a special
colorimetry.
For making accurate comparisons of intensity of
apparatus, called a colorimeter, must be used. The colorimeter of Duboscq is one of the best known and is selected for description.
ified
The colorimeter of Duboscq, as modshown in Fig. 181. The apparatus consists of an upright case, the front and sides of which are in one piece B, and hinged to the back. At the bottom of the case is a shelf S, containing
Duboscq's* Colorimeter.
by
Pellin, is
Circular of information.

two
circular openings,
465
C'.
above which
rest the
two cylinders C and
The latter are very carefully constructed, being closed at the bottom by disks of glass whose upper and lower surfaces are perfectly plane Two immersion rods of solid glass, T and T' the ends of parallel.
Fig. 181.
Fig. 182.
Duboscq's colorimeter.
which are also plane parallel are attached to movable slides in the back of the case and can be raised or lowered within the cylinders. The height of the lower surface of each rod above the bottom of its cylinder is indicated upon a scale, which by means of a vernier can be read to 0.1 mm. The colorimeter is illuminated by light from the reflector M, which from its opposite surfaces gives either bright or diffused
light according to the requirements of sensibility.
in Fig. 182, passes
upward through each
cylinder
The light, as shown and immersion rod to
466
the prisms
SUGAR ANALYSIS
and
P',
from which
it is
reflected
is
upwards into the
tele-
scope A.
The
field,
when the
telescope
focused, consists of a circle
F, divided into equal parts, exactly resembling the double field of a Daylight is to be preferred for illuminating the colorimpolariscope. eter although artificial white, or monochromatic, light may be used
according to requirement. In preparing the instrument for use, the mirror must be adjusted so that both halves of the field appear of exactly equal intensity. The sugar solution
which
is
to be tested
is
and the standard
solution, containing a
known percentage
placed in one cylinder of the same
sugar, in the other, both solutions having been previously treated under The similar conditions with alkali or other color-producing reagent.
door of the case is then closed and the rod immersed in the solution to be tested to some convenient scale division, as 100 mm., 50 mm., etc., at which point the color of its half of the field should be of suitable intensity for comparison.
of
The other rod
is
then immersed in the cylinder
standard solution, and lowered or raised until the two halves of the The heights of the immersion rods above field are of equal intensity.
the bottoms of the cylinders will then be inversely proportional to the depth of color and hence to the amount of sugar in solution. The calculation
If
is
made
as follows:
A= B = P= X=
=
the elevation of rod in standard solution, the elevation of rod in solution to be tested,
the per cent of sugar in standard solution, the per cent of sugar in solution to be tested,
then
AXP
B
c.c.
50 gms. of a glucose solution of unknown strength were made with up water, adding 5 c.c. of dilute NaOH solution (solution I). One gram of pure glucose was dissolved in water and the solution made up to 500 c.c. adding also 5 c.c. of the same NaOH solution (solution II). Both solutions were heated in a hot-water bath for the same length of time
Example.
to 500
and
after cooling
compared
in a
Duboscq
colorimeter.
the immersion rod in solution I was set at 100 mm., the immersion rod in solution II gave equal intensity to the field at 160.2 mm.
1
When
Then
VL
1-60 gms. of glucose in the 500
c.c. of
solution
I,
or 3.2
per cent in the original sample.
Johnson* has recommended heating with alkaline picric-acid

tion for the colorimetric determination of glucose.
*
solu-
Picric acid
is
reduced
Mon.
sclent., Ill, 13, 939.
467
by glucose and other sugars in alkaline solution to picramic acid, the deep red color of which is sharply developed by less than 0.01 per cent of sugar. As stable color standards Johnson recommends solutions of ferric acetate, or of ferric chloride and acetic acid, which have been prepared so as to match the color produced by a known weight of sugar under the conditions of the method.
Many
of the color reactions of sugars are affected
of organic or mineral impurities; the usefulness of colorimetric in estimating sugars is for this reason largely curtailed.
by the presence methods
Ehrlich's Colorimetric Method for Estimating Caramel. Ehrlich* has devised a colorimetric method for estimating caramel, in which the standard of comparison is saccharan. This dark-colored caramel subis produced by heating sucrose in a flask immersed in oil to about 200 C. under vacuum. The residue, after extracting with
stance
boiling
methyl alcohol,
is
9,
The saccharan, Ci 2 Hi 8
is
dissolved in water, filtered and evaporated. obtained as a dark-brown residue (about
20 per cent of the weight of sucrose) which is easily pulverized to an amorphous powder. One part of saccharan in 10,000 of water colors
the solution a deep brown, which
alkalies.
is
intensified
by the addition
of
Saccharan is not precipitated by lead sub-acetate solution, so if the latter is used for precipitating other coloring substances from solutions of sugars, molasses, etc., the percentage of saccharan in the neutralized filtrates may be estimated by comparison in a colorimeter with a solution containing a known weight of saccharan. The amount of saccharan multiplied by 5 indicates the approximate amount of sucrose destroyed by superheating during manufacture. Stammer's Colorimeter. Colorimeters are employed in technical
sugar analysis for grading sirups, for estimating the decolorizing power of bone black or other clarifying agent, and for many other purposes
in
which degree of
color,
and not determination
of color-producing sub-
For determinations of this kind colored plates, or stance, is desired. disks, of glass are usually employed as a standard of comparison, the results being expressed in units of an arbitrary color scale.
A
is
colorimeter which
is
used extensively
in the sugar industry is
The general principle of this apparatus (Fig. 183). the same as that of Duboscq. The liquid to be tested is placed in the cylinder a, which is closed by a glass plate at the bottom. The
that of
Stammer f
measuring tube
*
c,
also closed at the

59, 746.
bottom by a
glass plate, fits
Z. Ver.
Deut. Zuckerind.,
Proceedings, Seventh International Conp. 747.
gress of Applied
t
Chem., Sect., V, p. 92. " Stammer's " Zuckerf abrikation (1887),
468
loosely into a
SUGAR ANALYSIS
and can be' raised or lowered to any desired level. The is joined to c, the b, which is open at the bottom, two being moved in conjunction by a slide in the back of the instrument.
comparison tube
The
is illuminated by a rebottom, the light passing upward through b and c into the prisms in d which produce the same double-
colorimeter
flector at the
field effect as in
In
operating
the Duboscq apparatus. the colorimeter the
standard plate of colored glass is placed upon tube 6, which together with tube c is then raised or lowered until the intensity of shade for solution
and
color
plate is the same in both halves of the field. A millimeter scale upon the
back of the instrument marks the elevation of the measuring tube above the bottom of the cylinder, thus indicating the thickness of the column of liquid. Stammer gives a solution which matches the standard plate for a scale
reading of
1
mm., a
color value of 100.
The
color value of
any
liquid
is
found
of the
by dividing 100 by the reading

scale in millimeters.
In measuring the color of sugars, etc., a weighed amount of Stammer's colorimeter. Fig. 183. substance is dissolved in water, made up to a definite volume and, if the solution is not clear, filtered. The color value of the solution is then calculated either to the original amount of substance, or to a polarization of 100, according
molasses,
to requirement.
Example.
and
eter.
filtered.
Then
W=
The
20 gms. of a sugar, polarizing 92.4, were dissolved to 100 c.c. solution gave a reading of 15 mm. upon Stammer's colorim6.666 the color value of the solution.
:
The
:
color value
calculated to 100 parts sugar would be 20 6.666 latter calculated to 100 polarization would give 92.4
::
100
x
::
33.33.
:
The
36.07.
33.33
100
color value of the solution
For determining the decolorization produced by bone black the is taken before and after filtration. If the
original solution
METHODS
469
is too dark for reading in the colorimeter, it is diluted with water, in which case the filtered solution is also diluted to the same
density.
Example.
of 8
mm.,
or
An unfiltered sirup diluted to 10 degrees Brix gave a reading = 12.5 color units, using a Stammer colorimeter. The liquid,
after filtering
through bone black, and diluting to 10 degrees Brix gave a reador
ing of 40
mm.,
is
W=
10 K
2.5 color units.
The amount
80 per cent.
of color
removed by the
bone black
then -
9 ^
12.5
100
table of reciprocals (Appendix, Table 25) will be found convenient for converting the scale measurements of Stammer's colorimeter into color units.
DETERMINATION OF SUGARS BY WEIGHING AS HYDRAZONES AND OSAZONES
The varying
solubility of the different hydrazones
and osazones
of
sugars in presence of impurities, or of other similar derivatives, has prevented the general employment for quantitative purposes of this means of separating sugars. In certain cases, however, where the
hydrazone, or osazone, is characterized by great insolubility a fairly accurate determination of several of the sugars has been found possible. Determination of Arabinose as Diphenylhydrazone. According
arabinose is precipitated quantitatively by treating the sirupy solution of sugar with a slight excess of diphenylhydrazine. Sufficient alcohol is added to form a perfectly clear solution, and the
to
Neuberg
mixture heated to boiling for 30 minutes in a water bath in a flask connected with a reflux condenser. The solution is cooled, allowed to
stand for several hours and the white crystalline hydrazone filtered into a weighed Gooch crucible. After washing with a few cubic centimeters of cold alcohol, the crucible is dried in a water oven and weighed.
CsHioC^N N(-C 6 H 5 ) 2 = 0.4747. C 5 Hi 5 by multiplying by This method of analysis has been used by Neuberg for estimating arabinose in the urine and by Maurenbrecher and Tollens f for de-
The weight
of arabinose diphenylhydrazone,
,
is
calculated to arabinose,
termining arabinose in cacao.
The property Determination of Mannose as Phenylhydrazone. in forming with phenylhydrazine a very insoluble hydrazone, discovered by Fischer and Hirschberger,t has been used for the quantitative estimation of mannose. The precipitation, according to
of
mannose
Ber., 36, 2243.
f Ber., 39,
3578.
| Ber., 21, 1805.
470
SUGAR ANALYSIS
Bourquelot and Herissey,* is best accomplished by treating a 3 to 6 per cent solution of the sugar with an excess of phenylhydrazine acetate After standing 24 hours, the white at a temperature not above 10 C. crystalline hydrazone is filtered upon a weighed Gooch crucible, washed
The solucold water, dried in a water oven and weighed. the hydrazone is 0.04 gm. in 100 c.c. of solution, and the weight of precipitate should be corrected accordingly.
with a
little
bility of
mannose phenylhydrazone, CeH^C^^HCeHs, is calC 6Hi2 O 6 by multiplying by |f = f or 0.6666. The method is well adapted for determining mannose in presence of other sugars and has been employed by Pellet f for estimating small amounts of mannose in sugar-cane molasses.
The weight
of
culated to mannose,
to
Neuberg
Determination of Fructose as Methylphenylosazone. According { fructose may be determined with a fair approximation
by precipitating as its methylphenylosazone, CeHioO^^CHsCeH^. About 10 c.c. of the concentrated sugar solution are treated with a slight excess of methylphenylhydrazine, and sufficient alcohol added
If other sugars than fructose are present the to give a clear solution. solution is slightly warmed and allowed to stand 24 hours for the sepa-
any insoluble hydrazones of mannose, galactose, etc. After removing any precipitate by suction, the filtrate is treated with 4 c.c. of 50 per cent acetic acid, heated 5 to 10 minutes upon the water bath, and then set aside in the cold for 24 hours. The reddish-yellow crysration of
tals of
the osazone are filtered in a weighed
culated to fructose,
method
is
and calby multiplying by J|J = 0.4663. The only approximate as 10 per cent or more of the osazone
crucible
Gooch
C Hi2
6
6,
remains in solution.
ration has been
By
using a very cold freezing mixture the sepaquantitatively.
made almost
SIEBEN'S
METHOD FOR ESTIMATING FRUCTOSE
Sieben in 1884 proposed a method for determining fructose which based upon the destruction of this sugar when heated with dilute hydrochloric acid. The method was designed for estimating fructose in honey, sirups and other products which contain glucose. The latter
is
sugar, like other aldoses, is much less susceptible to the destructive action of acids, so that the difference in the reducing power of a solu*
Compt.
rend., 129, 339.
t Bull, assoc.
chim. sucr.
dist., 16,
1181;
18, 758.
t Ber., 35, 960.
Z. Ver. Deut. Zuckerind. (1884), 837, 865.

tion before
471
is
and
after treatment
by
Sieben's process
taken as the
equivalent of the fructose present.
In making the determination 100

contain about 2.5 gms.
250-c.c.
graduated flask
c.c. of the solution, which should total reducing sugars, are heated in a with 60 c.c of 6-normal hydrochloric acid
of
(36.47 bath.
prevent evapothen cooled and neutralized with 6-normal sodium hydroxide (40 X 6 = 240 gms. NaOH per liter), of which from 56 to 58 c.c. are usualty required. The contents of the flask are then made up to 250 c.c., filtered and the reducing sugars determined in 25 c.c. of the filtrate by Allihn's method. The reducing sugar thus found is
ration.
X 6 = 218.8 gms. HC1 per liter) for 3 hours in A funnel is placed in the neck of the flask to
The
solution
is
a boiling-water
calculated as glucose, and the difference in reducing sugar before after the acid treatment estimated as fructose.
and
According to Sieben only about 1.5 per cent of the total glucose is * destroyed under the conditions of his method. Herzf eld found, howWiechever, that the destruction of glucose may exceed 7 per cent.
mannf
also showed that the complete destruction of the fructose is not " always assured so that the results obtained by this method must be received with some caution." Dammullert found that the destructive
power
of the acid
depended largely upon the
ratio of glucose to fructose;
with mixtures of glucose and fructose in equal proportions only 1.28 per cent of glucose was destroyed, with pure glucose on the other hand the loss exceeded 28 per cent. Attempts to modify and improve the process so as to overcome these objections have not been wholly successful.
*
(1898), p. 54. Z. Ver. Deut. Zuckerind., 38, 751.
Wiechmann's "Sugar Analysis"
CHAPTER XVI
COMBINED METHODS AND THE ANALYSIS OF SUGAR MIXTURES
IN previous chapters upon polariscopic and chemical methods several instances were given of the application of certain processes to In the present chapter the problem of the analysis of sugar mixtures. in several sugars presence of one another will be taken up determining
in
somewhat
If
fuller detail.
of the specific rotations, copper-reducing powers or other properties of the different sugars in a mixture can be expressed by a sufficient number of equations, the problem of determining the
the
sum
percentage of each sugar in the mixture may be solved by simple algebraic analysis. By thus combining the results of several distinct
methods
it is
possible
by
indirect
means
to
sugar mixtures with a
fair
degree of accuracy.
make an analysis of many The combinations of
methods, which have been proposed for this purpose, are almost numberless and only a few examples will be chosen to illustrate the general The methods will be grouped for convenience under principle.
(1)
(3)
Combined Combined
polariscopic methods;
(2) Combined reduction methods; methods. reduction and polariscopic
COMBINED POLARISCOPIC METHODS

If two sugars, and B, exhibit a known variation in specific rotation under different conditions of polarization, then the percentages, x and y, of the two sugars may be determined by means of the following
equations:
ax
+ by =
lOOWz),
',
(1)
a'x+b'y=lW[a] D
in
(2)
which [oi\D and [U]D are the specific rotations of the mixture A + J5, a and a' the known specific rotations of sugar A and b and b the known specific rotations of sugar B, under the respective conditions of (1) and (2) By determining [O\D and [O\D the percentages x and y are readily calf
.
culated.
glucose
As an example of this method of analysis the determination of and fructose by polarization at 20 C. and 87 C., under the
472
COMBINED METHODS AND ANALYSIS OF SUGAR MIXTURES 473

conditions previously described (p. 296),
of glucose are
is
given.
If
the []*> and [a]%
the []g and cent fructose are
52.5 respectively, then of a mixture x cent containing per glucose and y per [a]g 92.5
+52.5 and
of fructose
and
52.5 x 52.5 x
92.5 y 52.5 y
= =
100[a]
100[a]g
By
determining the [a]" and []^ of the mixture the percentages of

is
glucose and fructose are readily calculated. Any other temperature, at which the [a]^ of each of the sugars
known,
may
of course
sults as thus calculated are of course only
be taken instead of 20 C. and 87 C. The reapproximate and require to
be corrected for the influence of concentration.

In addition to varying the temperature, changes of condition may be accomplished by making one polarization in neutral and the other in acid solution; or one polarization in water, and the other in some other solvent; or one polarization in the absence and the other in the presence of borax or other substance; in all of which changes of condition a definite known alteration in the polarizing power of one or both sugars must be produced. Obviously the greater the degree of
this
change in polarizing power, the
less will
be the influence of ex-
perimental errors.
COMBINED REDUCTION METHODS

If two sugars, A and B, exhibit a known variation in reducing power under different conditions of analysis, then the percentages x and y of the two sugars may be determined by means of the general
equations
ax
+ by =
100/2,
(1) (2)
a'x+b'y = lQOR',
in
which
and
a' the
known reducing powers
are the reducing powers of the mixture A f of sugar A, and b and b the
+ B, a and
known
re-
under the respective conditions of (1) and (2). R and By determining R', the percentages x and y are readily calculated. A good example of the application of the above formulae is given by Soxhlet's * well-known method for determining two sugars in mixture. A comparison of the reducing powers of different sugars upon Fehling's copper solution (Soxhlet's formula) and Sachsse's mercury solution was made by Soxhlet with the following results:
ducing powers of sugar B,
*
J. prakt.
Chem.
(1880), 21, 300;
Konig's
"
Untersuchung
"
(1898), 217.
474
SUGAR ANALYSIS
TABLE LXXXII
Showing Relative Reducing Power of Fehling's and Sachsse's Solutions
Sugar.

determining the values F and S of the mixture of sugars, the percentages x and y are readily calculated.
By
stants a,
In using the above, or other combined reduction methods, the conbj a' and &' should be determined empirically by the chemist
for the particular sugars
with which he
of
is
working.
As another example
combined reduction methods
may
be men-
tioned Kjeldahl's* process of determining the reducing power of the mixture of two sugars in both dilute and more concentrated solution,
using respectively 15 c.c. and 50 c.c. of mixed Fehling's solution according to the details of his reduction method (p. 424). The relative
differences in the copper-reducing
powers under the two conditions of
analysis are not sufficiently pronounced, however, to afford a reliable basis of calculation and the method has been generally condemned.
The use of combined polariscopic, or of combined reduction, methods alone for analyzing sugar mixtures has largely given place to the more accurate procedure of combining these two distinct physical and chemical
methods
in one.
COMBINED POLARISCOPIC AND REDUCTION METHODS

1.
ANALYSIS OF MIXTURES CONTAINING TWO SUGARS
calculation of the percentages of two sugars in mixture by comthe results of polarization and copper reduction was first atbining tempted by Neubauerf in 1877, and the principle of his indirect method
The
has been that of most subsequent modifications.
In the
earlier
methods
of this class the total reducing power of the mixture was determined as glucose, fructose or invert sugar, the percentage thus obtained being
taken as the total amount, or sum, of the sugars present. In the case of two sugars, A and B, the percentages x and y of each were expressed by the formula x + y = R in which R was the percentage of total reducing sugar determined as The results calculated by such a glucose, fructose or invert sugar. formula have, however, only an approximate value, as the difference in copper-reducing power of the two sugars A and B has not been taken
into account.
The error last mentioned has been largely obviated in the later methods of this class through the use of reduction factors (p. 421) by means of which the copper-reducing power of a sugar can be converted into the equivalent of any other reducing sugar which is selected as a standard of comparison. For the latter purpose glucose is usually
*
Z. analyt. Chem., 35, 345-347.
f Ber., 10, 827.
476
SUGAR ANALYSIS
of the reducing sugars and the selected, this being the most common one most easily obtained in a pure condition. It was shown upon p. 421 that the different monosaccharides bear a constant ratio to glucose for the same weight of reduced copper. This ratio was given for several sugars and was found by Allihn's method
to be 0.915 for fructose, 0.958 for invert sugar, 0.898 for galactose, 0.983 for xylose and 1.032 for arabinose.
For a solution containing a mixture of monosaccharides, the sum of the glucose equivalents of the individual sugars should equal the total reducing sugars estimated as glucose. This is shown in the following
* experiments by Browne, who mixed known weights of different sugars and compared the calculated glucose equivalents with the amount of glucose corresponding to the reduced copper obtained by Allihn's
method.
TABLE LXXXIII
Showing Glucose Equivalents of Mixed Reducing Sugars
Sugars.

the reducing ratio of a sugar remains the same whether
or with other monosaccharides.
it
occurs alone
General Formulae for Analysis of Sugar Mixtures. If the reducing ratio of sugar A to glucose is a, and of sugar B to glucose &, then in a mixture of x per cent A and y per cent B, the combined influence is represented by the equation:
ax
in
+ by
=R
(1)
1
which
If
is
the percentage of total sugars determined as glucose.
the relative polarizing power of sugar A be expressed by a and that of sugar B by j8, then in a mixture of x per cent A and y per cent B, the combined influence is represented by the equation:
ax
in
+ (3y
=P
(2)
which
By
the polarizing power of the mixture of sugars. combining equations (1) and (2) we obtain:
is
ab-ap
aR- aP
y
=-b^'
or
R- ax
<*>
the constants a, b, a and are known, the percentages x and two can be monosaccharides calculated very closely from the any y of total determined as glucose, and from percentage reducing sugar,
of
When
the polarizing power of, the mixture. In the following applications of Applications of the Method.* the preceding formulae to special problems of analysis, the polarizations were made upon a Ventzke-scale saccharimeter using the sucrose normal weight. The relative polarizing power of a sugar under these conditions is best expressed in terms of sucrose'and is found by dividing
rotation by the specific rotation of sucrose, or +66.5. In making up the various mixtures the sugars were weighed in a small stoppered flask. After adding the requisite amount of water the flask was reweighed and the percentage of each sugar in the solution calculated. After the sugars were dissolved, the solutions were
its specific
allowed to stand 24 hours before beginning the analysis, in order to remove all possibility of error through mutarotation.
Analysis of Mixtures of Fructose and Glucose. Reducing ratio of fructose to glucose = 0.915
Reducing
ratio of glucose to glucose
1.000
= =
a.
b.
* The applications of the method to the analysis of mixtures containing two sugars are taken from the paper by Browne upon "The Analysis of Sugar Mixtures," J. Am. Chem. Soc., 28, 439.
478
SUGAR ANALYSIS
C., 10 per cent solution) to sucrose
Polarizing ratio of fructose (20
-90.18
Polarizing ratio of glucose (10 per cent solution) to sucrose
By
previously given,
substituting the values for a, we obtain:
6,
a and
/3
in the general equations
Per cent fructose (F) Per cent glucose
7Q3
7?
_ P - =
0.381
Z.Uo
#-
0.481 P, at 20 C. (1)
(2)
=R-
0.915 F.
to the great susceptibility of fructose to variations in specific rotation through changes of temperature and concentration, the use of
Owing
a fixed polarization factor is only possible when the analyses are made under perfectly similar conditions. The values of the polarization
factor of fructose for different temperatures
and concentrations are
given below:
Tempera-

The following analyses were made known amounts of fructose and glucose:
Taken.
of seven mixtures containing
480
SUGAR ANALYSIS
= =
a.
b.
Analysis of Mixtures of Glucose and Galactose. Reducing ratio of glucose to glucose = 1.000 Reducing ratio of galactose to glucose = 0.898
Polarizing ratio of glucose (10 per cent solution) to sucrose 52.74
Polarizing ratio of galactose (20
+ 80 49 T66^~
-
21
we
substituting the values for a, obtain:
By
b, a.
and
/?
in the general equations,
Per cent glucose
G=
91
r>
f)
OQO
Per cent galactose
R-G
0.898
A ,no 0.498
2 43
-
R-
1.803 P, at 20 C. (3)
(4)
The
ture
specific rotation of galactose varies
somewhat with tempera-
however, being much less than for the polarization factor of values following concentrations were calculated and at different temperatures galactose from the general formula of Meissl.
and concentration, the
differences,
those of fructose.
The
Temperature. Degrees. C.
COMBINED METHODS AND ANALYSIS OF SUGAR MIXTURES
481
The average error in the above series of experiments is nearly four times that found in the separation of fructose and glucose. This was to be expected since, owing to the small difference in the specific rotations of glucose and galactose, the errors of observation are doubled; in the analysis of the fructose-glucose mixtures on the other hand the wide range in the specific rotation diminishes the experimental errors
one-half.
Analysis of Mixtures of Fructose and Galactose. Reducing ratio of fructose to glucose = 0.915
Reducing
ratio of galactose to glucose
0.898
= =
a.
6.
Polarizing ratio of fructose (20
90.18
+ 66.5
Polarizing ratio of galactose (20
1.356
a.
+ 80.49 + 66.5
By
equations
substituting the above values for a, we obtain:
o-j =I
1.21
b,
a and
0, in
the general
r>
r\
OQO
Per cent fructose (F) Per cent galactose
=0.521 fl- 0.386 P (20
C.).
(5)
p =-
A qi
c J?
- = 1.114 R
1.019 F.
(6)
The susceptibility of the specific rotations of both fructose and galactose to temperature variations necessitates a considerable correction if the polarizations are made much above or below 20 C. By using the polarization factors for fructose and galactose previously Thus given, formula (5) can be corrected for any desired temperature.
, ., Qn0 C. for 30 per cent fructose
-0.898 P 1.179# -
~o~ooi~
The following
amounts
analyses were
made
of four mixtures containing
known
of glucose
and
galactose.
Taken.
482
SUGAR ANALYSIS
Analysis of Mixtures of Fructose and Arabinose. Reducing ratio of fructose to glucose = 0.915
Reducing
ratio of arabinose to glucose
1.032
= =
a.
6.
Polarizing ratio of fructose (20 C., 10 per cent solution) to sucrose 90.18
Polarizing ratio of arabinose to sucrose
+ 66.5 + 104.5 + 66.5

}
= -1.356 =a.
1.571
j8.
By
equations,
substituting the above values for a, b we obtain:
a and
/3,
in the general
Per cent fructose (F)

Per cent arabinose
0*39
2.836
= 0.554#-0.364P(20C.).
(7)
0.915
1.032
0.969
R- 0.887 F.
(8)
Correction for changes in temperature
The following known amounts of

Taken.
is made as in the previous cases. analyses were made of two mixtures containing fructose and arabinose.

The
amounts
following analyses were made of four mixtures containing of xylose and arabinose.
known
Taken.
484
II.
SUGAR ANALYSIS
ANALYSIS OF MIXTURES CONTAINING THREE SUGARS
The indirect method of combining polarization and reducing power can also be applied, but with considerable limitations, to the analysis of mixtures containing three sugars. Methods Based upon a Determination of Total Sugars, Reducing
Power, and Polarization. The calculation of three sugars in a mixture is sometimes made (1) from a determination of the total sugars, as by drying or by densimetric means, (2) from the reducing power and (3) from the polarization. If three sugars A B and C constitute a mixture, and no other substances are present, the percentages x, y and z of each may be expressed
,
as follows:
x + y + z = T (total solids). + by + gz = R (reducing sugars as glucose) ax -f &y + 72 = P (polarization).
(1)
.
ax
(2) (3)
Having determined T, R and P, and knowing the reducing constants a, b, and g and polarizing constants a, /3, and 7 of the three sugars, the percentages x, y and z of each may be calculated in certain cases
with a
in
fair
degree of approximation.
calculations
It frequently happens,
however,
errors
making
by
this
method that small experimental
are enormously multiplied, so that the final results, even with mixtures of pure sugars, can be regarded as only very roughly approximate.
Analysis of a Mixture Containing Glucose, Galactose and Fructose. As an example of the limitations above mentioned the problem of analyzing a mixture containing x per cent glucose, y per cent galactose
and
per cent fructose is taken. By substituting the reducing and polarizing constants previously employed for these three sugars in the general equations (1), (2) and (3) we obtain:
z
x x whence,
z
0.793 x
+ + 1.21y0.898 y
+ y + z = T, + 0.915 z = R,
1.356 z
P, at 20 C.,
(1) (2) (3)
= = =
- 1.638 R - 0.401 P, at 20 C. per cent fructose = 1.957 T - 8.433 R 0.33 P, at 20 C. = cent T 8.175 per galactose = cent T z. per glucose y
errors in determining total solids or reducing sugars are magnified in the calculation of galactose over eight times.
It is seen that
any experimental
Example.
galactose
solution containing 6.46 per cent glucose, 8.22 per cent

results:
and 9.67 per cent fructose gave upon analysis the following

Total solids (T) by drying in vacuo 24.20 per cent; reducing sugars (R) as glucose 22.80 per cent; polarization (P for 26 gms. in 100 c.c., 200-mra. tube at
1.95 V. Substituting these values for T, R and P in the previous equations gives fructose 9.23 per cent; galactose, 6.21 per cent and glucose
20 C.)
8.76 per cent.
The relationships between experimental errors and the errors in calculated results in the above example are as follows:
486
SUGAR ANALYSIS
The relationship between experimental errors and the errors in calculated results in the above example are as follows:

Equation comes
If (1) of Allen,
expressed in
its
simplest decimal form, be(4)
m = 3.195 S + 4.642 K - 6.326 0.
the sample be polarized upon a saccharimeter, where the ratio of the scale reading for the normal weight to specific rotation will be as
the [O\D of sucrose (+66.5) is to 100, the factor for the saccharimeter reading of a normal weight would be for equation (4)
100: 66.5:: 3.195 :x
2.125.
Equation (1) of Allen modified for the polarization (P) of a sucrose normal weight upon a saccharimeter would then be:
m=
frequently calculated is done, a correction
in the solution
2.125
P + 4.642 K -
6.326 0.
In the analysis of starch-conversion products the total solids are
by means of the solution factor 3.86. When this must be introduced for the variations from 3.86 factors of the different ingredients. The solution factor
product has been placed
f actors f
of the mineral matter, or ash, in a conversion
at 8;* taking as the solution
dextrin (d), the values 3.83, 3.92 and 7 equation for total solids (T ) as calculated from the specific gravity the solution factor 3.86 (usually written T*.M) would be:
3.86
.
of glucose (0), maltose (m) and 4.21 respectively (p. 31), then the
by
3.86
3.86
3.86
Knowing the percentage

solids
of ash (a), the equation of Allen for organic
would be:
3.86
3.83
g
+ 3.92 m
,
3.86
,3.86, _ T ^4.21^
3.86
If
calculated
the reducing power be expressed in percentage of the solids as by the factor 3.86 (written Kê) then
3.86
.
3.86
3*3
In the same
*
32
by the
factor
way
if
the [O\D of the solids, as calculated
"Commercial Organic Analysis" (1901), Vol. I, 376. noted that the solution factors of glucose, maltose and dextrin increase in the order of their specific rotations. From this relationship Rolfe (J. Am. Chem. Soc., = 0.004023 - 0.000001329 (195 - []#), 19, 698) has derived a general equation S
Allen's
t It is
for calculating the specific gravity influence "of
when the value
for [a] D (obtained
and 1.045) is known. O'SuUivan solution factor.
any acid-hydrolyzed starch solution. by the factor 0.00386 between the densities 1.035 The value for S multiplied by 1000 will give of course the
488
3.86,
SUGAR ANALYSIS
be used instead of the []/> of the moist product, then, using the [<*]/> of glucose, maltose and dextrin
52.7 g
values of Allen for the
139.2
m+
198 d
= 100[a]
fl ,,,
trin
Several other methods of calculating maltose, glucose and dexhave been proposed. These are similar to that of Allen, except
that slightly different values are used for the polarizing and reducing constants.
It is seen that in the calculation of
maltose by Allen's method any
experimental errors in determining organic solids, reducing power or The value of the method in specific rotation are greatly multiplied.
the analysis of hydrolyzed starch products is still further diminished by the fact that no account is taken of isomaltose and of the various reversion products which are always present in materials of high conversion. Any reducing power and rotation due to other substances
than glucose, maltose and dextrin affect the accuracy of the method to a marked degree. Furthermore the dextrins of starch conversion are of a mixed character with different rotations and reducing powers, so that the selection of an initial dextrin of [O\D + 198 and negative reducing power is largely arbitrary. The percentages of glucose, maltose and dextrin in starch-conversion products, as calculated from determinations of organic solids, reducing
largely conventional quantities;
power and polarization
are, therefore,
may
the latter, when properly understood, serve, however, as a valuable means of comparison.
The methods of estimating three sugars in mixture which depend upon a determination of total sugars become largely valueless in the case of such products as molasses, fruit juices, honeys, etc., which contain varying amounts of organic and mineral salts, gums, and acids. With such materials a determination of dry substance, or of organic solids, gives too high a percentage of total sugars, and the results of the calculation may even lack the value of an approximation. It is, therefore,
sible in a
always the best plan to determine as mixture by direct means.
many
of the sugars as pos-
Methods of Calculating the Percentages of Three Sugars from the Combined Reducing Power and Polarization and the Direct Determination of One Sugar. ing x per cent A, y per cent
If in
and
:
a mixture of three sugars containper cent C, the percentage z of C
be determined by direct means, then x and y can be calculated by means of the following equations

ax
ax
+ by -f gz = R + fiy + yz = P
z
(total reducing sugars as glucose) ,
(polarization)
= Z (direct determination), ax -f by = R whence gZ, ax + (3y = P- yZ. Having determined R, P and Z and knowing the
reducing and polar-
izing constants of the three sugars, the percentages x and y calculated as described on page 477, for mixtures of two sugars. Several applications of the method will be described.
can be
Analysis of a Mixture Containing Glucose, Fructose and Sucrose. The sucrose is best determined by the methods of inversion, using
either the process of double polarization or that of copper reduction.
the polariscopic method be used, the inversion is best accomplished by means of invertase in order to eliminate the influence of the acid
If
upon the rotation of fructose. Knowing the percentage (S) of sucrose in a mixture containing x per cent glucose and y per cent fructose, and no other optically active or reducing substances, the percentages x and y can be calculated by means of the two equations
:
whence,
+ 0.915 y R (reducing sugars as glucose), olari ion of a sucrose normal 0.793 x - 1.356 y + S = P, 20 C. (P ) V / weight on a saccharimeter 0.793 R S-P + OAOr /1A = cent fructose = at 20 C.
x
per
=-^ Z.Oo
(1) (2)
per cent glucose
=R
0.915 y.
The determination of R will be a little too high, unless a correction is made for the slight reducing action of sucrose upon Fehling's soluThis correction can be made by using an empirical formula, tion. such as proposed by Browne for Allihn's method (p. 427), or by using
the special methods and tables for determining reducing sugars in
presence of sucrose.
Example.
The
solution
employed
in the previous
example
;
by the method
of inversion 16.27 per cent of sucrose (S}
(p. 485) gave substituting this and
the previous values, obtain:
R=
15.24,
and
P = + 17.05
16.27
at 25
C., in equation (1)
we
Fructose
_
=
0.793 (15.24)
15.24
-17.05
Z.Oo
pef cent
Glucose
0.915(5.43)
10.27 per cent.
These percentages agree more closely than in the previous example with the actual amounts of sugars taken, viz.: 5.43 per cent fructose, 10.02 per cent glucose and 16.16 per cent sucrose.
490
SUGAR ANALYSIS
Analysis of a Mixture Containing Glucose, Maltose and Dextrin. In addition to the method of Allen previously described, several processes have been devised for determining glucose, maltose and dextrin
in starch-conversion products,
which are based upon a direct deter-
mination of the dextrin.

Determination of Dextrin. Several methods have been proposed the direct estimation of dextrin in presence of other carbohydrates, but none of these has been found to give perfectly reliable
for
results.
The dextrin is sometimes precipitated from the sirupy solution by adding a large excess of hot 95 per cent alcohol, and stirring, after which the precipitate of dextrin is allowed to subside. The clear
solution when deposition is complete is decanted through a filter, the dextrin dissolved in a little water and again precipitated by adding alcohol as before. The process is repeated for a third time, after which
is washed into a platinum evaporating dish, and dried and weighed. The residue is then ignited and the weight of ash deducted from the weight of dried alcohol precipitate; the difference is estimated as dextrin. The difficulty with this method of estimation is
the precipitate
to precipitate all of the dextrin without occluding any of the glucose or maltose. The dextrin after repeated precipitations with alcohol
still reduces Fehling's solution; this may be due, however, to the presence of reducing maltodextrins as well as to the occlusion of
sugars.
Methods based upon a destruction
of reducing sugars
by fermenta-
tion or oxidation, and then calculating the residual polarizing power to dextrin have already been referred to (p. 301). The principal objection to the fermentation method is that most yeasts ferment or modify
dextrin to a greater or less degree so that the residual polarizing power does not represent that of the dextrins originally present. In Wiley's method (p. 306) of destroying reducing sugars by oxidation with alkaline
maltose
mercuric cyanide, it has been found that the polarizing power of is not completely destroyed and that the dextrins themselves
acid.
undergo partial oxidation to dextrinic
just described it is evident that the percentages of dextrin thus determined have only a nominal
Owing
to the limitations of the
methods
value.
Assuming that the residual polarizing power (P'), after destroying maltose and glucose, is due to an unchanged dextrin of [a] D 193, and and calling the [a] D of glucose (0) 53 and of maltose (ra) 138, sup-
posing the relative reducing powers of glucose and maltose to be 100
COMBINED METHODS AND ANALYSIS OF SUGAR MIXTURES

and 62*
starch-conversion product
g
respectively, the calculation of the percentages g, is made by Wiley f as follows:
491
m and d in a
(1)
+ 0.62m = R (total reducing sugars as glucose). 53 g + 138 m + 193 d = 100 P, (P = a D of product).

[
]
193 d
100 P', (P'

(2) gives
(2) (3)
aD
]
after destroying g
and m).
Subtracting 53 g
(3)
from
138
(1)
m=
100 (P
P').
(4) gives
(4)
Multiplying
by 53 and subtracting from 105.14m = 100 (P - P'} - 53 #,

1 C\C\ (
(5)
whence
m=
g
T^
.--L--.
~pf\
10 5 14
\Q 7?
0.951
(P-P')-
0.504/2.
(G)
= R -0.62m. =
(7)
^f'
[a
(8)
A sample of midzu ame (Japanese glucose) was analyzed by Example. the with following results: Wiley = 132.6 = P. [a\ D before fermentation
D
after fermentation
Total reducing sugars as glucose

Substituting these values in equations
= + 59.2 = P'. = 33.33 per cent =

and
R.
Maltose = 0.951(132.6 - 59.2) Glucose = 33.33 - 0.62(53.01)

Dextrin
(6), (7)
(8) gives:
0.504(33.33) = 53.01 per cent, 0.47 per cent,
10Q (59 2)
'
30.67 per cent.
.L
i/o
If
scale readings,
the sample be polarized upon a saccharimeter the factor for the P and P' of a sucrose normal weight would be for equa-
tion (6)
100:66.5:: 0.951
(6) of
0.632.
Equation Wiley modified for the polarizations of a sucrose normal weight upon a saccharimeter would then be = 0.632 (P - P') - 0.504 R.
Equation (8) of Wiley modified for calculating dextrin from the saccharimeter reading (P') of a sucrose normal weight would be P' 193
= &M d P
''
whenced =
^902'
The
cose,
on page 488 of the indirect method of estimating glumaltose and dextrin from organic solids, polarization and recriticisms
ducing power apply also to the method of calculation just described. * The ratio 62, or in decimal form 0.62, is strictly true only for O'Sullivan's method. The factor is less than this for other processes of copper reduction,
t
Wiley's "Agricultural Analysis" (1897), Vol. Ill, 288.
492
SUGAR ANALYSIS
to the
Owing
mixed character
of the dextrins in starch conversion
= 193, or of any other products, the selection of a dextrin of [O\D fixed value, as a basis of calculation is largely conventional. The presence of the unfermentable reducing sugar isomaltose and of opti-
cally-active reversion products also affects the accuracy of the method. Owing to these reasons, as well as to the general unreliability of the
methods
for estimating dextrin, the results of such calculations have frequently no absolute scientific value. The Applications of the Method to Other Sugar Mixtures.
general principle of combining the results of polariscopic and reduction methods with those of a direct determination in analyzing mixtures of
three sugars has been sufficiently indicated, and additional examples
need not be given.

limited extension.
of unone of the three sugars is a pentose or methylpentose, its percentage may be determined from the yield of furfural- or methylf urf ural-phloroglucide mannose may be determined from the yield of phenylhydrazone; lactose or galactose from the yield of mucic In combining the acid; raffinose by the method of inversion; etc. results of such direct determinations with those of polarization and reducing power, the chemist must consider in each case the limitations of the methods used and the extent to which experimental errors are
If
;
Such schemes of analysis obviously admit
multiplied in the calculation.
The
final test of
accuracy consists in applying the method to the
analysis of mixtures containing known amounts of the several sugars, and this verification should be made whenever possible.
III.
ANALYSIS OF MIXTURES CONTAINING FOUR SUGARS
Schemes of analysis have also been proposed for the analysis of mixtures containing four sugars, in which case, however, two of the members present must usually be determined by direct means. As a single illustration of such methods the following scheme is
given for analyzing a mixture containing g per cent glucose, / per cent fructose, s per cent sucrose and x per cent xylose.
0.793 S
= S (sucrose determined by method of inversion) = X (xylose determined from yield of furfural phloroglucide).
.
+ . + 0.283* = P (P f \ weight upon a sacchanmeter / g + 0.915/ + 0.983 x = R (total reducing sugars as glucose).
1.356/
olari
tion of a sucrose normalN
(2) (3) (4)
Substituting the known values of 0.793 0-1.356/ =
S and
P-Sfl-
X in
(1) and 0.283 X.
(2) gives:
(5)
0.983
(6)

Multiplying
(6)
by 0.793 and combining with
(5) gives:
'
per cent fructose
S
=
0.793
R-P-QA97X ~
"2082
per cent glucose
=R-
0.915 /
0.983
X.
application of such formulae as the above to the analysis of complicated mixtures of sugars usually involves, however, such a com-
The
bination and multiplication of experimental errors, that a scheme of

calculation, perfectly correct in theory,
is
shown
in practice to be
almost valueless.
It is scarcely necessary to remark that in working with unknown mixtures of sugars, each of the constituents present must be identified by careful qualitative tests before beginning the analysis.
For a description of other methods and schemes which have been proposed for analyzing different mixtures of sugars, the chemist is referred to Lipmann.*
Vol. I, 616-623; 894-899. See also Wiechmann's "Sugar Analysis" (1898), and the papers by Halenke and Moslinger (Z. analyt. Chem., 34, 263) and by Geelmuyden (Z. analyt. Chem., 48, 137) for other examples of calculation.
*
"Chemie der Zuckerarten,"
CHAPTER XVII
MISCELLANEOUS APPLICATIONS
THE present chapter will give several practical applications of the principles and methods previously described to a few selected problems of technical sugar analysis. large number of such applications have already been considered and a description of these will be passed
over.
The methods will be grouped under
three
main
divisions of prod-
ucts: (1) Sugar-factory products; (2) Starch-conversion products; (3)
Food products.
SUGAR-FACTORY PRODUCTS
In addition to the analytical methods, previously considered, a few definitions of common terms and several descriptions of illustrative
be given. For the application of methods to the technical works of Geerligs, Mittelstaedt, sugar-factory control, Pellet and Metillon and others should be consulted. Morse, Spencer,
will
commercial methods
The coefficient of purity of a juice, sirup, the molasses, sugar, etc., percentage of sucrose in the total solid matter of the product.' The term, which is also called " quotient of purity," "degree of purity," "purity" or "exponent," has been variCoefficient of Purity.
is
ously interpreted, and the chemist must distinguish carefully between the true and the apparent coefficient of purity.
The true coefficient of purity is the percentage of actual sucrose in the total solid matter as determined by the method of drying. The apparent coefficient of purity is usually taken as the ratio of the
direct polarization to 100 parts of apparent solids as calculated from the degrees Brix, or by other indirect means.
A sugar-cane molasses gave upon analysis Example. Total solids by actual drying ................... Total solids by degrees Brix .................... Total solids by refractometer ...................
the following result 75.10 per cent

77.10 per cent 74.20 per cent
Direct polarization ............................ 42.20 Sucrose by method of inversion .................. 45.70 per cent
True
Apparent
coefficient of purity
^x
10n
60.85. 54.73.
^~ X 100 =
(1)
494
Apparent
495
(2)
100
is
56.87.
Sometimes the true percentage of sucrose

purity in which case
used in calculating apparent
Apparent
= =
'
= =
59.27. 61.59.
(3)
'
Apparent
(4)
coefficient of purity of sugar-cane or sugar-beet juices loosely applied to the entire cane or beet.
The
is
often
Numerous
tables
and formulae have been calculated
for converting
apparent into true purities, but these can only be used classes of products for which they were designed.
upon the
is
special
Determination of Ash.
The determination
of ash
of great im-
portance in the technical analysis of sugar products. Several methods of the Association of Official Agricultural Chemists * are given. Direct Incineration. Heat from 5 to 10 gms. of sugar, molasses,
etc., in
the water
a platinum dish of from 50- to 100-c.c. capacity at 100 C. until is expelled, and then slowly over a flame until intumescence ceases. Then place the dish in a muffle and heat at low redness until a
is
white ash
Soluble
obtained.
and Insoluble Ash.
Add water
to the ash in the platinum
dish after weighing the total ash in the previous method, heat nearly to boiling, filter through ash-free filter paper and wash with hot water
until the filtrate
filter
and washings amount to about 60 c.c. Return the paper and contents to the platinum dish, carefully ignite and
weigh.
The
residue
is
the weight of insoluble ash.
The
difference
between insoluble and total ash gives the soluble ash.
Owing to the difficulty of obtaining a perfectly carbon-free ash and to the danger of expelling volatile salts during ignition Scheiblerf has recommended burning the sample in presence of sulphuric acid. Saturate the sample with sulphuric Ignition with Sulphuric Acid. Deduct acid, dry, ignite gently, then burn in a muffle at low redness.
one-tenth of the weight of the ash, then calculate the per cent. Instead of deducting one-tenth, to correct for the weight of combined sulphuric acid, Girard and Violette propose the deduction of
one-fifth.
When it is dePreparation of Ash for Quantitative Examination. sired to obtain a pure carbon-free ash for quantitative examination the
following
method should be used

* Bull. 107 (revised),
f
*
:
Carbonize the mass at a low heat,

Bur. of Chem., pp. 67 and 68.
4,
U. Stammer's Jahresbericht,
S.
221; 7, 267.
496
SUGAR ANALYSIS
the solution of soluble salts,
burn the residual mass to and evaporate to dryness whiteness, add and in a desiccator cool at 100 C., ignite gently, weigh. The percentage of ash deDetermination of Organic Matter.
dissolve the soluble salts with hot water,
ducted from the percentage of total solids gives the percentage of

organic matter.
The percentage of sucrose deducted Determination of Non-sugar. solids of total the from gives the percentage of non-sugars. percentage The percentage of ash deof Determination Organic Non-sugar. of the from ducted non-sugar gives the organic non-sugar. percentage This coefficient is found by dividing the perSaline Quotient.
centage of sucrose by the percentage of ash. The glucose ratio, or coefficient, represents the Glucose Ratio. of sucrose. It is found by multiplying the per100 of glucose per parts centage of reducing sugars by 100 and dividing by the percentage of
sucrose.
of the glucose ratio is of great importance in Any increase in this coefficient during clarification or evaporation indicates a partial inversion of sucrose. The term extraction has been Determination of Extraction.
The determination
sugar-house control.
given several meanings in consequence of which occasional confusions
and misunderstandings have arisen. In Louisiana and Cuba, extraction indicates the percentage of undiluted juice which is obtained from a given weight of cane. Thus, if
2000
Ibs. of
the juice has been diluted owing to saturation (i.e., spraying the ground cane with water before regrinding), its equivalent in undiluted juice must first be determined before mak-
100
cane give 1500 75 per cent.
Ibs. of
undiluted juice the extraction
is
If
ing the calculation. In the Hawaiian Islands, extraction means the percentage of sucrose in the cane that is obtained in the mixed juices and is calculated by the
,
lormuia
percent sucrose in mixed juice

:
per cent sucrose in cane

of
X weight of mixed juice X ^ cane X weight of

-.
1An luu.
Ibs.
2000 Ibs. of cane containing 15 per cent sucrose gave 2300 Example. mixed diluted juice which polarized 12.4. Then
1
9*300
i g lo
s/oTWT X ÛUU
10
95 07 P er cent extraction,
-
Determination of Acidity and Alkalinity of Sugar Products. The determination of the acidity and alkaHerzfeld's Method. of linity sugar products is at times a matter of considerable importance.
The Herzfeld
or
German
official
method
for determining the
acidity
497
and alkalinity of raw sugars following solutions are used:

in
is
selected for description.

is
The
One part of phenolphthalein (1) Phenolphthalein. 30 parts of neutral 90 per cent alcohol.
(2)
dissolved
Neutral
Water.
Ten
liters of freshly
boiled distilled water
are treated with 5
the phenolphthalein solution and sufficient dilute alkali (see under 4) added to produce a permanent pink tinge. The water should be prepared several hours before use, but should
c.c. of
not be used after one or two days as the indicator loses

bility.
(3)
is
its sensi-
prepared,
(4)
is
A n/280 sulphuric acid solution Standard Sulphuric Acid. 1 c.c. of which is equivalent to 0.0001 gm. CaO.
Standard
Sodium Hydroxide.
1
solution
prepared,
c.c.
of
A n/280 sodium-hydroxide which exactly neutralizes 1 c.c. of the

the pink tinge of the
standard acid.
Ten grams
water
*
of the sugar are dissolved in 100 c.c. of the neutral

If
in a porcelain evaporating dish.

is
neutral water
discharged the sugar is acid and the acidity is measured by noting the volume of standard alkali necessary to restore the If the pink tinge of the neutral water is reddened the original color.
sugar is alkaline and the alkalinity is measured by noting the volume If the of standard acid necessary to bring back the original tint. end-point of the titration is over-run, the solution is titrated back
with acid or alkali as the case may be. The acidity or alkalinity of the sugar is then expressed in the equivalent percentage of CaO. Thus 10 gms. of a sugar requiring 30 c.c. of standard acid for neutralization
would have an
Juice.
alkalinity of 0.03 per cent
CaO.
true normal juice is the mixed juice as it It is impossible to actually exists in the tissues of the cane or beet. obtain this true normal juice by any method of pressing or milling for
Normal
The
reasons explained on page 232, so that its composition and percentage must be calculated by indirect means. In cane-sugar factories it is often customary to call the undiluted juice of the first mill the normal
juice
and
to
make
all
calculations
upon
this basis.
more
correct
practice is to determine the degrees Brix and polarizations of the different mill juices and then by means of empirical factors, established for the conditions of each factory, to calculate the approximate percentage
and composition
* (Int.
of the
normal
juice.
With dark sugars a

Sugar
larger
J. 13, 305), in
volume of the neutral water must be taken. Cross a modification of Herzf eld's method, employs 200 c.c. of
neutral water.
498
SUGAR ANALYSIS
The Dutch standard consists of a series of Dutch Standard. samples of cane sugar ranging in color from a very dark No. 7 to" an almost white No. 25. These samples are put up each year in sealed bottles by two firms in Holland, under the direction of the Netherlands
Trading Society, and are sent to different parts of the world as color standards for classifying sugars in the assessment of duty. The relation between color and composition is such a loose one that the Dutch standard has purely an arbitrary value. Calculation of Rendement.* - - The rendement is the yield of pure The crystallized sucrose which can be obtained from a raw product. various formulae, employed in its calculation, subtract from the polarization, or sucrose content, of the product a certain quantity which is taken to represent the melassigenic influence of the ash or other nonof the most common methods of calculation is that first Monnier in France in 1863; Monnier assumed that 1 part proposed by
sugars.
One
of mineral impurities
and so calculated the
prevented the crystallization of 5 parts of sucrose, yield of crystallizable sugar by subtracting 5 times the percentage of ash from the polarization of the raw product. This method of calculation is very largely used in the valuation of raw
beet sugars.
For cane sugars the following formula

(5
is
often used:
.
Rendement = Polarization
per cent ash
per cent invert sugar)
Monnier's formula for calculating rendement is used, however, more in other countries than in France itself. The method most used in France at present is to subtract from the polarization 4 times the
percentage of ash and twice the percentage of invert sugar; from this remainder 1.5 per cent additional is then deducted as the loss in refinIn 1893 the German Refiners' Association introduced a method ing.
for calculating rendement of total non-sugars by 2J
" was found, however, to be less satnon-sugar yield " " than the ash and a return was made to the old isfactory yield method of Monnier. The number of methods used by different assozation.
This
"
which consisted in multiplying the percentage and subtracting the product from the polari-
and factories for calculating rendement is almost unlimited. Determination of Crystal Content. The calculation of rendement by formula is unsatisfactory for the reason that the variations in melassigenic influence of the non-sugars are not considered. A direct
ciations
determination of the sucrose crystals in a raw sugar has, therefore, been proposed as a better means of determining the refining yield.
For a very full discussion of methods for calculating the refining value, " net analysis," or rendement of raw sugars see Mittelstaedt's "Technical Calculations for Sugar Works."
*
499
The different methods for determining sugar ^Method of Payen. content are all modifications of the early process of Payen,* which consisted in washing the adhering sirup from the crystals of raw sugar by means of 88 per cent alcohol, saturated with sugar and containing
50 c.c. of strong acetic acid per liter. The object of the acid was to break up saccharates and promote the solution of calcium carbonate and other mineral matter. The method of Payen was displaced in
1871 by the following modification of Scheibler.f
Method for Determining Crystal Content. The four washin have Scheibler's method the used following composition: ing liquids (1) 85 per cent alcohol containing 50 c.c. of strong acetic acid per liter is saturated by shaking with an excess of powdered sucrose. (2) 92 per cent alcohol saturated with sucrose as (1). (3) 96 per cent alcohol saturated with sucrose as (1).
Scheibler's
(4)
mixture containing 2 volumes of absolute alcohol and
1 vol-
ume
of ether.
Stock solutions (1), (2) and (3) are preserved in large double-neck bottles (Fig. 184), which are filled, as is also the siphon tube $, with lumps of loaf sugar. The tube T contains calcium chloride for preventing absorption of moisture from the air. The solutions should not be exposed to wide changes in temperature. In making the determination a half-normal weight of the ground sample of sugar is placed in a 50-c.c. graduated flask F, which is closed with a two-hole stopper. One hole of the latter is fitted with the inlet
7, through which the washing liquids are added, and the other with the outlet tube 0, through which they are withdrawn. The tube extends to the bottom of the flask and at its lower enlarged end is fitted
tube
with a
The large bottle B, which receives the spent filtering plug of felt. washing liquids, is connected by the opening of its stopper to the outlet tube of the sugar flask and by the side opening to a suction pump. The alcohol-ether solution (4) is first run into the flask F, using about 2 volumes to 1 volume of sugar. After standing 10 minutes, with occasional shaking, the liquid is sucked off into B. The alcoholether removes moisture from the sugar and at the same time precipiThe tates sucrose from the film of sirup adhering to the crystals. sugar is then treated in exactly the same way with solutions (3) and (2) the latter remove the traces of alcohol and ether left from (4) and prepare the sugar for the action of solution (1) which accomplishes the
;
Dingier 's Poly tech. Journal (1846), 100, 127.
t Full descriptions of Scheibler's
experiments are given in Stammer's Jahres-
bericht, Vols. 12
and 13 (1872 and 1873).
500
SUGAR ANALYSIS
Solution (1) is next added, using the same chief part of the washing. After before. as shaking 10 to 15 minutes the spent liquor proportions
is
withdrawn, and a second portion of (1) added; the process is continued with (1) until the washings become colorless. The sugar is
Fig. 184.
Scheibler's apparatus for determining crystal content of
raw
sugars.
then treated with solutions

as
(2), (3)
and
(4) in
the order named.
After
removing gently warmed, while a strong current of air is drawn through to remove the last traces of alcohol and ether. The connections are then removed from F, any particles of sugar adhering to the tube 0, or plug of felt, washed into the flask, and sufficient water added to dissolve the contents. A few
is
much
of (4) as possible, the flask
501
drops of lead reagent are added, and the volume completed to 50 c.c. The solution is then filtered and polarized; the saccharimeter reading
gives the percentage of sucrose crystals in the sugar. The method of Scheibler for determining crystal content has not given satisfactory results and is at present but little used. It has been
found that a considerable precipitation of sucrose may take place from adhering wash liquors especially upon contact with the alcohol-ether. The precipitation of sucrose from the molasses in the sugar is also
objectionable, especially when and amount of such molasses.
it is
desired to calculate the composition
In order to reKoydl's Method for Determining Crystal Content. duce the above-named errors and simplify the manipulation, Koydl* has recently modified the Payen-Scheibler method as follows The following five washing liquids are used: (1) 82 per cent (by weight) alcohol containing 50 c.c. concentrated
:
acetic acid per liter.

(2)
86 per cent (by weight) alcohol containing 25
c.c.
concentrated
acetic acid per liter.
91 per cent (by weight) alcohol. 96 per cent (by weight) alcohol. All of the above solutions are saturated with sucrose in the cold, and kept over lump sugar in stock bottles.
(3)
(4)
(5)
Common
absolute alcohol.
In making the determination 50 gms. of sugar are weighed into a beaker of ordinary form, 18 cm. high; 250 c.c. of solution (1) are measured into a wash bottle from which a sufficient quantity is added to the beaker until the sugar is covered about 1 cm. deep. After well
mixing, the solution is poured through a weighed filter paper (16 cm. diameter) in a covered funnel. The process is repeated several times, the sugar being finally transferred to the filter and washed with solution
(1) until
the 250
c.c.
are used.
When the
(4) are
filter
has drained completely,
poured in successive portions upon the sugar, each liquid being allowed to filter off before adding the one following. The sugar is then washed with 100 c.c. of (5) taking
c.c.
50
of solutions (2), (3)
and
When the alcohol has filtered well the edges of the paper. are dried in an oven and then its contents completely, the paper and of weighed. The weight product multiplied by two gives the crystal
care to
wash
content of the sugar. Koydl's method has been found to give results which are approximately quantitative, when the requirements of uniform temperature,
*
Oester. Ungar. Z. Zuckerind., (1906), 277.'
502
SUGAR ANALYSIS
saturation of solutions and other details are carefully maintained. With variations from these requirements a considerable error may result
gum
from solution or precipitation of sucrose. A certain amount of and mineral matter is always precipitated from the adhering
molasses
by the
alcoholic solutions;
the final crystals
when
dried
polarize from 99.4 to 99.8 and contain about 0.2 per cent organic nonsugar and 0.15 per cent ash.
Results of analyses of several beet sugars giving the composition,
rendement (polarization less 5 times ash) and crystal content by Koydl's method are given in Table LXXXIV which is taken from results by
Ehrlich.*
TABLE
LXXXIV
No.
503
There are other modifications of Payen's method for determining crystal content, but none equal in practicability to those of Scheibler and Koydl.* Such methods have found their chief value not in the work of routine but in providing a control upon other processes. In many European refineries the crystal content of raw sugars, massecuites, etc., is determined by washing a large sample of the product in a laboratory centrifugal (Fig. 185) with a saturated sugar The results obtained by a practical test of this kind are often sirup. found to have more value than those obtained by any modification of the Payen method. In order to avoid the Method of Herzfeld and Zimmermann. error due to the use of alcoholic washing fluids, Herzfeld and Zim-
have recently devised a method for determining crystal content, by which the raw sugar is simply shaken and washed with a saturated aqueous sucrose solution. The latter is always prepared just before use by weighing out 500 to 600 gms. of water in a strong glass-stoppered flask and adding the exact amount of sucrose to produce saturation at the laboratory temperature, which should be as
mermann
near 20 C. as possible. The grams of sucrose necessary for saturating 100 gms. of water at temperatures between 15 and 35 C. are given in the following table:
Laboratory
temperature.
504
its
SUGAR ANALYSIS
f
;
lower end by the rubber plug S 200 c.c. of the saturated sucrose
ber stopper
solution are then added, the rubS is inserted and the
whole shaken vigorously until all molasses adhering to the sugar The crystals has been dissolved.
vessel
should not be handled
with the bare hands, which might warm the solution and dissolve
some of the crystals. The vessel A is then attached
by the rubber stopper B,

removing the plug
ing cup C.
after
The
is
$', to the filteropen bottom of

filter
the latter
plate h
}
closed with a
pad
of
upon which rests a thin felt /, and above the latter

g.
a disk of wire gauze and gauze are previously cleaned, dried and weighed.
The
felt
The cup C
ter-flask
is
then
fil-
attached to the
D and the
a
stopper S replaced by a stopper containing
small
tube.
capillary
Suction
is
then
applied and the contents of A are
gently discharged into C. The inner

surface
of
is
then washed with a little sugar solution from

all
until
crystals
are Fig. 186.
Herzfeld and Zimmermann's apparatus for deter-
about mining crystal content of raw sugars. removed; 50 c.c. of sugar solution are sufficient. The cup, without sucking
off
all
505
the sirup, is then placed in a small centrifugal and whirled for 5 minutes, in the first minute at 2000, in the second minute at 2500, and for the remaining time at 2700 revolutions per minute. The cup is then removed and its contents are discharged into a weighing
bottle
by inverting and gently pushing the bottom
any
crystals left adhering to the walls of the
removed.
dried in
The vacuum
sugar, felt and gauze at a final temperature of 105

felt
plate with a rod, cup being also carefully are weighed, and then to 110
C.
After
and gauze the loss by drying is calculated to sugar-sirup by multiplying by the ratio of water to sirup for
deducting the weight of
the temperature of saturation. The weight of sirup deducted from the weight of sugar after centrifuging gives the weight of crystals. The method of calculation is illustrated by the following example:
A
21
saturated sugar solution was prepared for a laboratory temperature of

is 1
:
C.; the ratio of water to sirup for this temperature
3.024.
Weight Weight Weight
of
raw sugar taken
of sugar after centrifuging of sugar after drying
= = = = = = =
50.00 gins. 48.62 gms.

48.01 gms.
0.61 gms. 1.84 gms.
Difference due to water of sirup 3.024 Adhering sirup = 0.61
Weight
of crystals
48.62
1.84
Crystal content of sugar
46.78 gms. 93.56 per cent.
Results of analyses of several samples of raw beet sugar by the above method are quoted from the work of Herzfeld and Zimmer-
mann.
506
SUGAR ANALYSIS
as yet been generally from its deserves The process but recognition simplicity. tested, should be subjected to a careful control according to individual
The Herzfeld-Zimmermann method has not
technical requirements. Calculation of Composition
and Purity
of
Molasses
in
Raw
Sugars. contained in
knowledge raw sugars
of the composition and purity of the molasses is often desired. The determination is made
indirectly by subtracting the sucrose of the crystals from that of the raw sugar and calculating the remaining ingredients as due to molasses.
The purity
of the molasses in sugar
Number
2 of the previous table

Per cent.
would be calculated as follows:

raw sugar = 100.00 content of raw sugar Crystal
Dry substance
Difference
of
3.20
= = = = =
=
96.80
=80.60
16.20
= Dry =
substance of molasses in raw sugar

in
Polarization of
raw sugar Polarization due to crystals

Difference
89.90
79.79
10.11
raw sugar
80.60
X X
0.99
Polarization due to molasses in

of molasses in
raw sugar
'
Apparent purity
raw sugar
1A on lo.zu
100
62.4
STARCH PRODUCTS
Several methods Polariscopic Methods for Determining Starch. have been devised for estimating starch from the specific rotation, after conversion into the soluble form. The following methods* have been used. Solution of Starch by Heating Under Pressure. From 2 to 3 gms. of material are heated in a 100-c.c. flask with 80 to 90 c.c. of water until a uniform gelatinization of the starch has been obtained. The flask is then placed in an autoclave (Fig. 175) and heated 3 to 5 hours
at 2 to 3 atmospheres' pressure. After cooling, the clear solution is to 100 c.c., filtered and polarized. The soluble starch thus obtained is without action upon Fehling's solution; its rotation is
made up
[<*]D
= +196.5
to
+197.
starch in the 100
c.c. of
solution
Using the value +196.5, the weight of is calculated from the angular rotation
[O\D
a in the 200-mm. tube by means of the formula
' c rrj
whence
grams starch
100 a
(+
196.5)
Solution of Starch by Means of Hydrochloric Acid. Five grams of the starch-containing material are rubbed with 20 c.c. of concentrated
*
Wiley's
"
Agricultural Analysis
"
(1897), Vol. Ill, 205.
507
hydrochloric acid of 1.17 sp. gr. for about 10 minutes. When the solution has cleared, the volume is completed to 200 c.c., and the liquid filtered and polarized. The soluble starch as thus prepared has a rotation of = 196.3 to +196.7. Using the mean value of +196.5, the grams [a]o of starch in 100 c.c. of solution are calculated as in the previous method.
With impure starch-containing materials, neither of the above polariscopic methods has the accuracy of the diastase method described on page 440.
Calculating the Apparent Composition of Starch-conversion Prod* ucts from the Specific Rotation. Brown, Morris and Millar have
100
100
s
in
"
>,co
a &
20
.
3.0
MD
190
170
150
130
110
[aj
Specific Rotation
Fig. 187.
Showing
relation of specific rotation to composition of acid-hydrolyzed
starch products.
shown that
tion exists
between the
in starch products of diastase conversion a constant relaspecific rotation and copper-reducing power of
the total solids.
Rolfe and Defrenf have also shown that in starch
products of acid conversion the solids of same specific rotation have " always the same reducing power irrespective of the source of the .starch, the nature or amount of the hydrolyzing acid, or the temperaIt is, ture conditions, these influencing the rate of hydrolysis only." the relationship betherefore, possible to express by means of a curve
tween
specific rotation
and copper-reducing power, or between
either
of these constants and the apparent percentages of glucose, maltose and dextrin, calculated by means of such formulae as are used in Allen's
method (p. 486). Upon this principle Rolfe has prepared the diagram shown in Fig. 187, which gives the percentages of dextrose, maltose and dextrin in the dry substance of starch-conversion products cor*
J.
Chem.
Soc., 71, 115.
t J.
Am. Chem.
"
Rolfe,
Soc., 18, 869;
The
"
Polariscope
(1905), p. 197.
508
SUGAR ANALYSIS
responding to the values of [a] D for dry substance (as determined by the solution factor 3.86) between +195 for dextrin and +53 for
glucose.
value, for example, of
[a] D
=+
100 for the dry substance
(cal-
culated from the density of an approximately 10 per cent solution at 15.5 C. by the solution factor 3.86) of an acid-conversion product
would correspond to an apparent composition of dry substance of 10 per cent dextrin, 40 per cent maltose and 50 per cent glucose. The apparent percentages as thus determined are useful for purposes of comparison and valuation but must not be mistaken for
absolute percentages for reasons already given. As Rolfe is careful to " there are comparatively few commercial products pure enough to permit of their constitution being determined in this simple manner."
state
The following method has Analysis of Commercial Dextrins. been used by the United States Bureau of Chemistry in testing dextrins for the National Bureau of Printing and Engraving. The method is a
modification
F.
by Browne and Bryan Lippmann.f

Specific Rotation.
of a
scheme
of analysis proposed
by
Transfer 10 gms. of the finely divided sample
and after solution in about 50 c.c. of cold water add 5 c.c. of alumina cream and make up the contents to 100 c.c., thoroughly shake and filter. Polarize the filtrate in a 200-mm. tube, using any form of polariscope or saccharimeter. It is important that a 6 per cent solution of bichromate of potash in a 3-mm. tube be used as a
to a 100-c.c. flask,
light filter.
tion
is
In using a Ventzke-scale saccharimeter, the OA ao vx TT in which found by the formula []/> = -
specific rota-
~^
V=
Ventzke
reading.
Dissolve 100 gms. of dextrin in 200 c.c. of cold water mortar or porcelain dish, and determine the visby rubbing up cosity of the solution by any of the standard forms of viscosimeter.
Viscosity.
in a
Comparative results should always be made by the same instrument and under similar conditions of temperature a uniform length of time
;
should also elapse after making up the solution before taking the visThe viscosity should be determined again on the same solucosity.
tion after standing 24 hours,
Moisture.
and also after 48 hours. Determine by drying from 2 to 5 gms. of sample
for
4 hours at a temperature of 105 C. Absolute constancy in weight cannot be attained on account of the slow decomposition of the dextrin.
" Seventh Int. Cong. App. Proc., Sec. V, Z. t Spiritusind., 25, 304, 307, 316, 317.
*
Chem.," London, 1909,
p. 337.
Ash.
509
Five to 10 gins, of the sample are weighed in a tared platinum dish and burned over a flame at a low heat. The ash should not
be heated to fusion, otherwise loss from volatilization will occur. If a filtered hot-water solution of the dextrin Soluble Starch. with iodine solution, soluble starch is indicated. blue reaction a gives
lots of dextrin, 10 gms. each, into 100-c.c. flasks, add 50 c.c. water to each and after all soluble matter is dissolved make up the contents of the one flask with cold water at 100 c.c., shake and filter. Evaporate 20 c.c. of the solution (2 gms.) to dryness and dry for 4 hours at 105, as under determination of moisture. Weight
Weigh two
of cold
of residue, less ash
matter.
after
on incineration, equals cold-water soluble organic Heat the contents of the second flask to boiling, and then The weight of hotcooling make up to 100 c.c., shake and filter.
water soluble organic matter in 20 c.c. of solution is determined as beHot- water soluble organic less cold-water soluble organic gives fore.
the soluble starch.
If the residue insoluble in hot water shows Unconverted Starch. under the microscope grains, which are colored blue with iodine, unconverted starch is present. To determine the percentage, collect the residue insoluble in hot water on a filter, wash until free from soluble matter, and determine the starch by the usual methods. Determine in an aliquot of the cold-water Reducing Sugars.
soluble
by the method
Dextrin.
,
.
ducmg
of Allihn, the results being expressed as glucose. Subtract the specific rotation of the dextrin due to re, (52.5 X per cent reducing & sugar as glucose) A from the sugars
JLUU
original specific rotation of the sample. Multiply the remainder by 100 and divide by 186 ([a]n of dextrin *) to obtain the calculated per-
centage of dextrin in the sample. Undertermined Solubles. The per cent of cold-water soluble organic matter less calculated percentage of dextrin gives the percentage of
II
undetermined solubles. In Table LXXXV eight analyses of commercial dextrins by the above method are given. It is noted that with a decrease in specific rotation there is a uniform decrease in viscosity and in the calculated percentage of dextrin,
186 of dextrin is given by Schultze (J. prakt. Chem., 28, 327). [<x] D considerably lower than the figures +195 to +205, which have been reported by other authorities for carefully purified dextrins. The value +186 is used only as a commercial standard of comparison, and the percentages of dextrin
This
is
The
thus calculated have no strict scientific value.
510
and a uniform increase
SUGAR ANALYSIS
in reducing sugars
and undetermined matter.
A large
percentage of reducing sugars indicates over-dextrinization, and accompanying this there is always a formation of other decomposition
products.
TABLE
LXXXV
Giving Analyses of Commercial Dextrins
of the starch
511
*
the
marked
show following analyses by Jago difference in composition between extracts made by cold-
by the
diastase.
The
water and warm-water mashing.
TABLE
LXXXVI
Constituent.
512
expressed as degrees
SUGAR ANALYSIS
Lintner, represent the
copper-reducing power
produced by the action of a measured volume of the extract upon a

solution of soluble starch at 21
Soluble-starch Solution.
C. for
hour.
is
containing 2 gms. of soluble starch (prepared as described on page 577) in 100 c.c. In determining the diastatic power of malt, or flour, Procedure.
solution
made
25 gms. of the finely ground material are digested with 500 c.c. of water at room temperature for 5 hours. The solution is then filtered
until perfectly clear.
metal rack and 10 c.c. of the solublethe first tube 0.1 c.c. of the filtered starch solution added to each. tube 0.2 c.c., and so on, the tenth to the second malt solution is added, shaken and then placed for 1 c.c. The tubes are tube receiving 1.0
Ten
test tubes are placed in a
To
hour in a water bath kept at 21 C., 5 c.c. of mixed Fehling's solution are then added to each tube and the .rack is placed in a boiling-water bath for 10 minutes. The rack is then removed and, after the precipitates of cuprous oxide have settled, the two tubes are selected in which the copper is all reduced and in which some of it still remains in solution, as is shown by the absence or presence of blue color, or by means of the ferrocyanide test. The amount of malt solution just necessary to reduce the 5 c.c. of Fehling's solution is between the amounts added to these two tubes; the corrected amount is then assumed to be midway between these limits, or the value of the second decimal estimated from the depth of blue color in the tube where reduction is incomplete. A malt is given a diastatic value of 100 on Lintner 's scale when
0.1 c.c. of the filtered 5 per cent extract just reduces 5 c.c. of Fehling's
solution under the above conditions.
If 0.25 c.c. of
malt solution were
required to reduce the 5 c.c. of Fehling's solution then the diastatic

'
power
of the
malt would be
100
(J.dd
40 degrees Lintner.
slight
correction remains to be
tion
is
made
for the reducing sugars in the
malt solu-
any reducing power found by taking 5 c.c. of Fehling's solution, 10 c.c. of starch solution and 10 c.c. of water and heating to boiling. The malt solution is then added from a burette until the blue color is just discharged. If 7 c.c. of malt solution were used then there would be a correction of
=
and
for
of the soluble starch.
This correction
1.4 degrees Lintner to
be subtracted from the value pre-
viously found. In the case of evaporated malt extracts of high diastatic power a 1 per cent or 0.5 per cent solution of the extract is used, the values thus
513
obtained being multiplied by 5 or 10 to obtain the true degrees Lintner for a 5 per cent solution.
Lintner 's
Method
as Applied to Diastases.
In determining the
activity of diastase preparations Lintner* uses the method described for malt, the only difference being that the results are expressed in
terms of a diastase of which 0.12 mg. produces sufficient sugar to reduce the 5 c.c. of Fehling's solution. In making the test, from 50 to 100 mgs. of the diastase to be tested are dissolved in 4 to 5 c.c. of water and then made up to 100 c.c. or 200 c.c. according to the supposed
strength of the enzyme.
If under the conditions described for the malt method 0.2 mg. of a diastase was required to produce sufficient sugar to reduce the 5 c.c. of Fehling's solution, then its diastatic power 12 X 100 = 60 degrees Lintner (diastase scale) would be
.
It should
(
be noted that 100 degrees diastase are over 40 times
0^2
J as powerful as 100 degrees malt
upon Lintner's
100
scale.
Sykes Method.
and Mitchell's In the method
Gravimetric
of
Modification
f
1 c.c.
of
Lintner's
Sykes and Mitchell
c.c. of
2 per
cent soluble-starch solution are treated with
malt extract (prepared as in Lintner's method) 50 c.c. of Fehling's solution are then added and the liquid heated quickly to 98 C., when it is placed in a boiling- water bath for 7 minutes. The reduced copper is then determined, the weight of which divided by 0.438 (the grams of copper in 50 c.c. Fehling's solution) and multiplied
of the 5 per cent at 21 C. for 1 hour;
by 100
gives the diastatic
The
results are said to
power in degrees of the Lintner scale. compare well with those obtained by Lintner's
method.
closer degree of estimation
gravimetric method for determining diastatic power permits a than is possible by the original Lintner
Slight errors of estimation by the volumetric method cause considerable differences in the final results, when only small volumes
process. of diastase solution are taken.
Thus between
0.1 c.c.
and 0.15
c.c.
the
degrees Lintner (malt) will vary between 100 and Determination of Diastatic Power of Commercial
66.6.
Amylases,
Method
In studying methods Sherman, Kendall and ClarkJ for determining the diastatic power of commercial pancreatin, Sherman, Kendall and Clark found that the conditions of temperature and
of
*
J.
prakt.
Chem.
[2],
34, 378;
36, 481.
t Analyst, 21, 122.

J J.
Am. Chem.
Soc., 32,
107a
514
SUGAR ANALYSIS
activation under which an amylase normally works should be incorporated in the method. These authorities also showed that the
amount of reduced copper does not stand in simple proportion to diastatic power, different diastatic values being obtained when different weights
of
enzyme were taken.
These differences are due to the influence
of
rate of conversion; if the velocity of the reaction be considered, however, the same diastatic power is derived from the weight of reduced copper for any weight of
variations in the concentration of starch
upon the
enzyme. The following gravimetric method was used. The enzyme may be dissolved in pure water if its power Enzyme. If it is to stand, it should be dissolved in is to be tested immediately. water containing 4 c.c. of fiftieth-molar disodium phosphate per 100 c.c. The test should be made within an hour in any case. The amount of enzyme to be weighed out will depend entirely on its strength. These will doubtless differ with the different Activating Agents. amylases. For pancreatic amylase acting on 2 per cent starch, add 300 mgs. sodium chloride and 7 c.c. of fiftieth-molar disodium phosphate
per 100 c.c. (final volume) of reaction mixture. Procedure. Prepare 400 c.c. of 2 per cent soluble-starch solution and the enzyme solution of such a strength that 1 c.c. will contain from
mg. of enzyme. By means of a 1 c.c. Mohr's pipette, accurately calibrated in hundredths, measure into four 200 c.c. Erlenmeyer flasks such volumes of the solution as will contain 0.2, 0.5, 0.8 and 1.0 100 c.c. of the starch solution, premg. of enzyme, respectively.
0.4 to 1.0
Now
viously warmed to 40 C. is poured into each flask and the digestion allowed to proceed for 30 minutes, the temperature being accurately maintained at 40 C. At the expiration of the 30 minutes, stop the re-
by mixing at once with 50 c.c. of Fehling's solution and immerse the flask in a large bath of boiling water for 15 minutes. See that the water of the bath is kept boiling and that it stands above the level of the contents of any of the flasks. At the end of this heating filter quickly and determine the reduced copper by any accurate method.
action quickly
Correct the weight of reduced copper or cuprous oxide found for by subtracting from it the a blank test in which the starch solution is weight
the reducing power of the soluble starch " " obtained in
treated directly with the Fehling reagent. Of the four determinations thus corrected, select the highest weight of cuprous oxide which does
not exceed 300 mgs. and find the corresponding value of in the followThis value of ing table. divided by the milligrams of substance gives the diastatic power of the enzyme upon Sherman's scale.
Values for
Cuprous
oxide.
515
K from Cuprous Oxide Found
516
SUGAR ANALYSIS
MISCELLANEOUS FOOD PRODUCTS
detection and estimation of sugars in food products are made according to the physical and chemical methods previously described.
The
Such methods are often valueless, however,
for
many
purposes of the
food chemist, frequently desires to know more about the origin of his the sugars in product than about their nature or exact amount.
who
whether maple or the cane. Neither from does an derived its sucrose was sugar maple in and dextrin a determine whether invert the estimation of honey sugar these have been gathered by the bee or have been added as an adulterIn the solution of such problems as these the food chemist ation.
polarization of
his decision upon reactions and estimations of other ingredients than sugar, such, for example, as the amount of matter precipitated by lead subacetate or by alcohol, the composition of the ash and
sugar, for example, will not determine
must base
organic non-sugars, miscroscopical examination, etc. Such determinations lie strictly outside the province of sugar analysis and only a few For a fuller typical applications of such methods will be considered.
description of such processes the chemist is referred to the special works upon food analysis by Leach, Wiley, Allen, Blythe, Konig and
others.
DETERMINATION OF LEAD-SUBACETATE PRECIPITATE
The determination
of the
amount
of lead-subacetate precipitate
is
frequently used as a means of distinguishing pure maple sugars and The method sirups from those which are adulterated with cane sugar. is based upon the presence in maple products, and the absence in cane
sugars, of salts of malic acid subacetate.
which gives a copious precipitate with lead
tate.
Method for Measuring the Volume of Lead PrecipiThe apparatus consists of a glass tube and Apparatus. holder as shown in Fig. 188. The tube and holder weigh about 50 gms., and should be so constructed that when fitted together the bottom of
Hortvet's*
a
the tube will be exactly even with the lower surface of the holder. In set, each couple, tube and holder should be made to balance one an-
other.
When placed in the centrifuge there should be as nearly as possible a balanced load carried at the circumference of the wheel.
Determination.
of sugar,
Introduce into the tube 5
c.c.
of sirup or 5 gms.
add 10 c.c. of water and dissolve. Add 0.5 c.c. (10 drops) of alumina cream (prepared as directed on page 223) and 1.5 c.c. of lead sub*
J.
Anthem.
Soc., 26, 1523.
acetate and shake thoroughly. Allow the mixture to stand from 45 to 60
517
minutes, occasionally giving the tube a twisting motion to facilitate the Place the settling of the precipitate. tube with its holder in the centrifugal machine and run 6 minutes under the conditions given below.
If
any material adheres to the
sides
of the wider portion, remove it by means of a small wire provided with a loop at the end. Return the tube
to the centrifuge and run 6 minutes longer at the same rate. Note the
volume
to 0.01
of the precipitate, estimating c.c. as closely as possible.
Run
a blank, using water and the re-
agents named above, and correct for same. In the case of a sirup the re-
reduced to the 5-gm. basis by dividing by the specific gravity of the

sult
is
Fig.
188. Hortvet's apparatus for measuring volume of lead precipitate.
sample.
The centrifuge used in this method has a radius of 18.5 cm. and is run at a speed of 1,600 revolutions per minute. The velocity at the circumference of the wheel is computed in centimeters per second.
Calling
M (mass) unity in the formula F = Mv
2
>
the numerical expres-
sion for F, the centrifugal force, becomes 519,363. By measuring the radius (r) for any given machine
for F, the
and substituting
numerical constant determination above, the velocity for a machine given may be determined by the following formula, v = VFr. Given the velocity in centimeters per second, the required number of
revolutions per second or per minute can be computed. The volume of lead precipitate, as determined above, was found
by
Hortvet to vary from 0.94 c.c. to 1.82 c.c. for pure maple sirups, and from 1.18 c.c. to 4.41 c.c. for pure maple sugars. Adulterated maple sirups gave from 0.23 c.c. to 0.95 c.c. and adulterated maple sugars from
0.10
c.c.
to 1.40
c.c.
Winton's* Method for Determining Precipitated Lead (Lead Number). Weigh 25 gms. of the material (or 26 gms. if a portion of
*
J.
Am. Chem.
Soc., 28, 1204.
518
the filtrate
is
SUGAR ANALYSIS
to be used for polarization)
and transfer by means

c.c.
of
boiled water into a 100-c.c. flask.
Add
25
of standard lead-sub-
acetate solution,
and
10
filter
c.c.,
to the mark, shake, allow to stand at least 3 hours a through dry filter. From the clear filtrate pipette off dilute to 50 c.c., add a moderate excess of sulphuric acid and
fill
100
c.c.
of 95 per cent alcohol.
crucible,
wash with 95 per cent
Let stand over night, filter on a Gooch alcohol, dry at a moderate heat, ignite
at low redness for 3 minutes, taking care to avoid the reducing cone of Calculate the amount of lead in the precipithe flame, cool and weigh. tate using the factor 0.6831, subtract this from the amount of lead in 2.5 c.c. of the standard solution, multiply the remainder by 100 and divide by 2.5, thus obtaining the lead number.
The standard lead-subacetate
is
volume of lead-subacetate reagent of 1.25 water, and filtering if not perfectly clear.
prepared by diluting a measured sp. gr. with 4 volumes of
The lead number, as determined above, was found by Winton and Kreider to vary from 1.19 to 1.66 for pure maple sirups, and from 1.83 to 2,48 for pure maple sugar. Adulterated maple sirups gave lead numbers ranging from 0.02 to 0.92.
Raw cane sugars Limitations of the Lead-precipitate Methods. and hence contain all such as are made without clarification (especially
the organic salts of the juice) may give amounts of lead precipitate which are as great as those obtained with pure maple products. Doo* little and Seeker give, for example, the following comparison between
a Venezuelan muscovado sugar (" Melada ") and a pure Vermont
maple sugar.
TABLE LXXXVII
Determination.
519
ANALYSIS OF ASH AS A MEANS OF DETERMINING THE ORIGIN OF SUGARS
One
sugar
is
of the
most valuable methods
to determine the composition of
of ascertaining the source of a its ash. The mineral con-
and sugar cane show very pronounced differences, and, notwithstanding the influences of
stituents of the juice of the maple, sugar beet
clarification
and crystallization, certain of these constituents find their raw sugar in sufficient quantities to afford a valuable way basis of opinion. Sugar-beet juice, for example, in distinction from that of the cane and maple, contains considerable potassium nitrate and perceptible quantities of the latter are usually present in raw beet
into the
Even the higher grades of beet sugar will frequently respond to sugar. delicate tests for nitrates and this has been used as one means of distinguishing beet from cane sugar. As an example of the application of the ash-analysis method the following results by Doolittle and Seeker* upon the muscovado and
maple sugar
comparison.
of
Table
LXXXVII
are given.
Average determinations
made by Jones
f upon the ash of pure maple sugars are also added for
TABLE LXXXVIII
Analysis of the Ash of Muscovado and Maple Sugar
Determination.
520
SUGAR ANALYSIS
potassium oxide, calcium
It is seen that in certain constituents, as
oxide,
and sulphur
show very soluble and water-insoluble ash and of the alkalinities of the are valuable aids in forming an opinion as to the origin of a The ash for such determinations should be prepared according method described for quantitative examination (page 495)
.
trioxide, the ashes of the muscovado and maple sugars pronounced differences. The determinations of waterlatter
sugar. to the
DETERMINATION OF ALCOHOL PRECIPITATE

of substance precipitated by strong used in alcohol examining sugar-containing products. frequently The materials which are precipitated by alcohol may consist of mineral In many or organic salts, pectin, dextrin, dextran and other gums.
is
The determination of the amount
cases a qualitative examination of the alcohol precipitate throws considerable light upon the origin of the product.
Determination of Alcoholic Precipitate in Fruit Products. Method Chemists* Evaporate 100 c.c. of a 20 per cent solution of the fruit product to 20 c.c.; add slowly and with constant stirring 200 c.c. of 95 per cent alcohol and allow the mixture to stand over night. Filter and wash with 80 per cent alcohol by volume. Wash this precipitate off the filter paper with hot water
of the Association of Official Agricultural into a platinum dish, evaporate to dryness, dry at 100
C. for several
called alcohol
hours and weigh;

residue as ash.
precipitate.
then burn
loss in
off
the organic matter and weigh the

is
The
weight upon ignition
The ash should be
largely lime
and not more than 5 per cent

If it is larger
of the
than this some of the salts of the organic acids have been brought down. Titrate the water-soluble portion of this ash with tenth-normal acid, as any potassium bitartrate precipitated by the alcohol can thus be estimated. The general appearance of the alcohol precipitate is one of the best
total weight of the alcohol precipitate.
indications as to the presence of glucose and dextrin. Upon the addition of alcohol to a pure fruit product a flocculent precipitate is formed with no turbidity, while in the presence of glucose a white tur-
bidity appears at once precipitate forms.
upon adding the
alcohol,
and a thick
gummy
amount
When the
of sugar is
quantity of
gum
or dextrin
is
large, a considerable
sometimes occluded in the alcohol precipitate. This is especially the case with honey, for determining the dextrin in which
Browne has modified the

*
alcohol precipitate
method
Chem.,
as follows.
p. 80.
Determination
Method.
of
521
*
with 4 mark.
c.c.
Browne's Precipitate in Honey. Eight grams of honey are transferred to a 100-c.c. flask of water and sufficient absolute alcohol to complete to the
Alcohol
A little care is required to effect the complete removal of the honey from the weighing dish without using more than 4 c.c. of water.
transference
is
The
best
made by decanting
as
much
as possible of the
honey into the flask, then adding 2 c.c. of water to the dish to take up any adhering honey and again decanting. By using 1 c.c. more of the water in two successive washings and adding a few cubic
liquefied
centimeters of the absolute alcohol each time before decanting, the
honey can be completely transferred without the necessity of using more water than the 4 c.c. Absolute alcohol is used finally to rinse out the dish and is then added to the flask with continual agitation After shaking thoroughly until the volume is completed to 100 c.c.
the flask
sides
is
and bottom and the supernatant

is
allowed to stand until the dextrin has settled out upon the liquid has become perfectly clear
(usually in 24 hours). The clear solution

cipitated residue
washed with 10
then decanted through a filter and the prec.c. of cold 95 per cent alcohol to re-
move adhering liquid, the washings being also poured through the filter. The residue adhering to the flask and the particles which may
have been caught upon the filter are dissolved in a little boiling diswater and washed into a weighed platinum dish. The contents of the latter are then evaporated and dried in a water oven to constancy in weight. Should the amount of precipitate be considerable, it is necessary to dry upon sand in vacuo at 70 C.
tilled
latter is redissolved in
After determining the weight of the dried alcohol precipitate the water and made to a definite volume. The
following dilutions are employed in
making up the
1.0-1.5
solutions:
2.0-2.5
Weight of precipitate in grams
0.0-0.5
0.5-1.0
1.5-2,0
2.5-3.0
Volume
of solution
in cubic
centimeters
50
100
150
200
250
300
are then determined in aliquots from the filtered soluThe total tion of alcohol precipitate both before and after inversion.
precipitate less invert sugar and sucrose gives the per cent of dextrin. f While this method of estimating dextrin in honeys gives much more accurate results than the direct weighing of the alcohol pre*
The sugars
Bull. 110, U. S. Bur. of
Chem.,
p. 19.
is
gives a large amount of alcohol precipitate, it found best to take only 4 gms. of honey for analysis; in other respects the
t
With honeydew honey, which

of procedure
is
method
the same.
522
cipitate, it
SUGAR ANALYSIS
can not be said in any
it is
of the honey, although

close approximation.
way to give the true dextrin content believed that the figures obtained are a
amount
of dextrin always escapes preis taken of those
small
cipitation with alcohol;
furthermore no account
may be occluded in the alcohol precipitate other than the sugars, and no correction is made for the copper-reducing power of the honey dextrin itself. This latter factor, though apingredients which
parently very small, might prove to be of some importance if much dextrin were present. Notwithstanding these limitations, however, the percentage of dextrin as determined by the method described has been found to have a decided value, especially when it is wished to
compare honeys
of different origins.
The percentages of dextrin in different American honeys, as determined by the above method, is given in the following table of comThe honeys are position, which is taken from the work of Browne.
arranged in order of their dextrin content.
TABLE
LXXXIX
BuU.
110,
Giving Composition of American Honeys.
U. S. Bur. of Chem.
Kind
of honey.
'MISCELLANEOUS APPLICATIONS
523
strongly dextrorotatory ([]/> varies from about +115 to +160) and the presence of much honey dew may 'cause honey to polarize to the
right.
If commercial glucose is suspected, honeydew dextrins may be distinguished from those of starch conversion by dissolving the alcohol precipitate in a little water and adding a few cubic centimeters of
iodine solution; a red color, due to erythrodextrin, indicates the presence of commercial glucose.
PAET
II
THE OCCURRENCE, METHODS OF PREPARATION, PROPERTIES AND PRINCIPAL REACTIONS OF THE SUGARS AND ALLIED
DERIVATIVES
CHAPTER
THE
XVIII
CLASSIFICATION OF THE SUGARS AND THEIR FORMATION IN NATURE

sugars, of which some thirty or more have been isolated from animal substances, are among the most widely distributed, and plant in nature. compounds organic
The
trisaccharides
sugars proper, including the monosaccharides, disaccharides, and tetrasaccharides, are colorless, odorless, crystalline
substances, usually of sweet taste, and for the most part easily soluble in water. The more complex anhydride condensation products of the
sugars, the polysaccharides, are usually amorphous compounds of The entire group of saccharides, the little or no solubility in water. so-called carbohydrates, constitute approximately three-fourths of the
dry matter of the plant world. A simple sugar, or monosaccharide, may The Simple Sugars. be defined as an aldehyde or ketone alcohol of the aliphatic series, the molecule of which contains one carbonyl and one or more alcohol
groups, one of the latter being always adjacent to the carbonyl group.
H-C-O-H
All sugars contain, therefore,
C =O
as a characteristic group
upon the presence of which nearly all of the chemical properties of the sugars depend. The simplest possible sugar according to the above is
glycol aldehyde.
H H-C-O-H.
H-C=O
Sugars containing the aldehyde group are termed aldoses
-C-O-H H-C = (H-C-O-H

I
characteristic aldose group
and those containing the ketone group ketoses
H-C-O-H
C=0 -icharacteristic ketose
group
527
528
SUGAR ANALYSIS
According to the number of their carbon atoms the monosaccharides are divided into dioses (C 2 H 4 2 ), trioses (C 3 H 6 3 ), tetroses (C 4 H8 4 ), pentoses (C 5 Hi 5 ), hexoses (C 6 Hi 2 O 6 ), heptoses (C 7 Hi40 7 ), octoses (C8 Hi 6 O 6 ) and nonoses (C 9 Hi 8 O 9 ). There are also substituted monosaccharides in which one or more hydrogen atoms of a diose, triose,
tetrose, etc., are replaced
by a methyl group,
3 2
as, for
2 2
example, methyl3
diose
(CH C H O2 ),
3 2 3
3 5 3 ),
dimethyldiose
(CH C H
7
CH ),
7 3
methyltriose
(CH 3 C H
methyltetrose
(CH C H
3 4
04), methylpentose
(CH 3 C5 H9
etc.
5 ),
methylhexose
cules of
(CH 3 C6 Hn0
6 ),
The Compound Sugars.
methylheptose By the condensation of

3
(CH C Hi O
7 ),
2,
3 or 4 mole-
the monosaccharides, the disaccharides, trisaccharides and In such condensations one molecule less tetrasaccharides are formed.
of water
is
eliminated than the

6 2
number
of reacting sugars, thus
H O = CtfHÔn 2C Hi 6 2H2 O = Ci8H 32 Oi6 3C 6 Hi O 4C Hi 2 6 - 3H2 O = C24 H 42 2

2 2 6 6
(disaccharide).
(trisaccharide).
(tetrasaccharide).
The Polysaccharides. By the condensation of an indefinite number of molecules of the monosaccharides the polysaccharides are
formed.
In such condensations one molecule
less of
water
is
probably
eliminated than the total
number
of reacting sugar molecules, as, for
example
nC H O - (n - 1) H O = Pentose - (n - 1) H O = nC H
5 10
(C 5 H 8 O 4 ) n H 2 0.
Pentosan.
12
(C 6 H 10 O 5 )nH 2 O.
Hexosan.
Hexose
The quantity n
polysaccharides
is
may
usually so large, however, that the formulae of the be taken as simply (C 5 8 O 4 ) n (C 6 Hi O 5 )n, etc.,
without sensible error.
The term carbohydrate is a general one which Carbohydrates. In its original frequently applied to the entire group of saccharides. sense it was applied only to such saccharide substances as contain six, or a multiple of six, carbon atoms and have their hydrogen and oxygen in
is
the proportion to form water.

as simple
compounds hydrate. Thus

:
of carbon
Such substances were regarded loosely and water, and hence the name carbo12 6 2
= 6 C + 6 H O. H On = 12 C + 11 H O. Raffinose, Ci H O = 18 C + 16 H 0. Cellulose, (C H O )n = n (6 C + 5 H,O).

Glucose,
CH
6
Sucrose, Ci 2
22
2
2
32
16
10
writers, although
This original meaning of -carbohydrate is still retained by some it was proved long ago that the term can no longer be
CLASSIFICATION OF SUGARS
AND FORMATION
IN
NATURE
529
taken in its former literal sense. A large number of sugars contain less than six, or a fractional multiple of six, carbon atoms, and there are also many sugars whose hydrogen and oxygen atoms have a different ratio than in water, such, for example, as the methylpentoses, C 6 Hi 2 05. Alcohol and Acid Derivatives of Sugars. The term carbohydrate is very often extended to include the alcohol and acid derivatives of the simple sugars. While this extension of meaning is not approved
of
by all chemists, a knowledge of these compounds so closely allied to the sugars is indispensable. The monosaccharides, as aldehydes, stand midway between the alcohols and acids. They are easily reduced to
other.
the former on the one hand and readily oxidized to the latter on the Such reactions take place continually in the chemical processes of plant
and animal
life,
and
also occur in the industrial opera-
tions of sugar factories, distilleries, etc. proper understanding of this relationship is, therefore, of great importance. The following table,
which gives a
clear.
classification of the alcohols, sugars
and acids
of differ-
ent monosaccharides, will
The
the mutual relationship of these more members, which are found in nature either free or in a
make
polysaccharide form, are printed in heavy type.
TABLE
Showing Group Relationships
Group.
XC
Sugars and Acids
of Alcohols,
530
SUGAR ANALYSIS
TABLE
XC
(Continued)
of Alcohols,
Showing Group Relationships

Group.
Sugars and Acids
AND FORMATION
IN NATURE
531
External Compensation. Van't Optical Inactivity of Sugars. Hoff* called attention to the important fact that when a compound with an asymmetric carbon atom is produced in the vegetable or animal organism it is found in most cases to possess optical activity. When, however, such a compound is formed synthetically, from an inactive Van't Hoff showed that in the substance, optical activity is wanting. latter case inactivity was due to the two opposite isomers being produced in exactly equal amounts, whereas in nature only one of these isomers is formed. Thus the fructose produced in nature is levorotatory; the fructose made synthetically from acrolein dibromide is optically inactive, and consists of equal proportions of left-rotating and If the synthetic fructose be fermented, however, right-rotating sugar.
the left-rotating sugar polarize to the right.
is
destroyed,
when the unfermented isomer
will
In addition to the above case of external Internal Compensation. compensation between two asymmetric carbon compounds, there is also the case of optical inactivity through internal compensation between two opposite symmetrical halves of the molecule. Thus mesotartaric acid can be given either of the following configurations:
COOH
COOH
HOCH
I
HOCH
COOH
HCOH HCOH
I
COOH.
These apparently opposite forms are identical, however, for one configuration can be brought into coincidence with the other by rotating through an angle of 180. The two C atoms printed in heavy type
are each asymmetric, yet the compound is inactive, since the optical In such effect of the one is counterbalanced by that of the other.
cases of internal compensation the molecule can be divided by a plane of symmetry (indicated above by the dotted line) into two opposite
halves,
which are mirror images of each other.
Optical inactivity through internal compensation cannot exist with the sugars or their monobasic acids; it is common, however, with the sugar alcohols and dibasic acids. Mesoerythrite, adonite, xylite, dulcite,
mucic and allomucic
acids, ribo-
and xylotrioxyglutaric acids are
other examples.
Since every optiof or isomer, equal but exactly cally active substance has an antipode, is of considerable isomers of such opposite rotation, the nomenclature * " Chemistry in Space," Oxford (1891), p. 38.
Nomenclature
of Optically
Opposite Isomers.
532
SUGAR ANALYSIS
importance. In only a few cases, as with fucose and rhodeose, where the compounds were named before their antipodal nature was discovered, have wholly distinct names been given to the members of an
The optical antipodes of known sugars were first synopposite pair. thesized by Fischer* who adopted the plan of distinguishing such compounds by means of the letters d and 1. These symbols, which
= dexter, right / = Icevus, primarily refer to the character of rotation (d indicate Fischer to used were synthetic relationships rather than by left) directions of rotation. Fischer, starting with the common dextrorota; ,
tory sugars, glucose and galactose, gave them the symbols d-, and their All sugars which could be derived opposite isomers the symbols 1-. from these sugars synthetically were grouped in the corresponding d1- class. Ordinary fructose, or levulose, which though levorotatory can be synthesized from d-glucose, was, therefore, named d-fructose. Ordinary xylose is dextrorotatory but was called 1-xylose by Fischer,f because its first discovered synthetic relationship connected it with
and
1-glucose.
Salkowski and Neuberg afterwards found that ordinary
xylose could be derived from d-glucose through d-glucuronic acid. As Fischer remarks, had this latter relationship been discovered first,
he would have named the sugar d-xylose. Such a nomenclature has obviously more historic than scientific value, and various improvements have been proposed by Maquenne,J Rosanoff, and others. The original system of Fischer, however, is still the one without change in the present volume.
most used and
is
retained
Racemic mixtures,
i.e.,
mixtures of optical antipodes in equal pro-
The combined symbol d, 1-, introduced by Fischer, expresses the nature of such a combination more The letter i-, clearly than the symbol i-, which has also been used. however, is sometimes employed to designate iso-, and sometimes to specify a compound which is inactive through internal compensation, the latter use being the one followed in the present work. The Formation of Carbohydrates in Nature. The carbohydrates are formed primarily only in the plant world, the proximate constituents of their formation being carbon dioxide and water. The combination of
portions, are necessarily inactive.
||
these
a process called assimilation chlorophyll-bearing tissue of the leaves.

*
is
The carbon dioxide

"
effected only in the green (3 vol-
of
Maquenne's Les Sucres." J. Am. Chem. Soc., 28, 114. For a very complete treatment of the subject of assimilation and of the origin carbohydrates in plants the reader is referred to Czapek's " Biochemie der
J t Ber., 40, 102.
II
Ber., 23, 370; 40, 102.
Pflanzen," Jena, 1905, Vol.
I,
pp. 188-583.
AND FORMATION
IN
NATURE
533
umes of which occur in 10,000 volumes of air) enters the leaf through the breathing pores and there unites with the water which has been drawn up through the roots from the soil. The combination takes
place with the liberation of one volume of oxygen for each volume of carbon dioxide assimilated. The process is thus the opposite of respiration and combustion, as is illustrated by the following equations:
Respiration and Combustion ........
C Hi O
6 2
Sugar
C0 + 6 H 0. +6O = 6 <a
2
2
+ oxygen =
2
+
6
water.
Assimilation ...................... 6
CO + 6 H O = C Hi O +
2
6 2
2.
dioxide
water
sugar
+ oxygen.
Assimilation in building up sugar thus plays an important part in purifying the atmosphere and in keeping a balance in the economy of nature.
Photosynthesis. Assimilation takes place only by daylight and is most active in the bright sunshine. The chlorophyll grains constitute the mechanism by which the energy of the light waves is transformed into
chemical work;*
it
has been observed that light in passing through the

is
green coloring matter of chlorophyll
wave
length,
and
this
phenomenon
plays,
changed from shorter into longer no doubt, an important part
in the process of assimilation.
hidden in obscurity.
is
The many intermediate steps in the process of assimilation are still The most widely accepted view, that of Baeyer,f
is
that formaldehyde
the
2
first
2
product formed,
C0 + H
The
fact that
is
= CH
2.
formaldehyde is found in green leaves only in the smallest explained by assuming that it immediately undergoes a condensation to form a hexose carbohydrate,
traces
CH O = C H
2
6
12
6.
The condensation by Loew
of formaldehyde to a mixture of hexose
sugars has been advanced as an argument in support of this theory. Opinions differ widely as to the nature of the carbohydrate which
Many plant-physiologists and chemists be to glucose, from which all the other carproduct Others believe starch to be the are derived. afterwards bohydrates
is first
formed
in assimilation.
consider the
first
.
* The fact that the light from the sky is more or less polarized has given rise to the hypothesis that the energy of such polarized sunlight produces the optical The hypothesis has found activity of the sugars which are formed by assimilation.
no
scientific support.
t
Ber., 3, 63 (1870).
534
first
SUGAR ANALYSIS
carbohydrate
formed and others sucrose. Glucose, fructose, and starch have all been detected in the leaves of maltose, sucrose, ease but the with which the different sugars in nature pass into plants, one another by condensation or hydrolysis makes it difficult to say
whether
It
is
this or that sugar
well established that the starch of the leaf

If
of photosynthesis.
primary or secondary origin. is one of the products the leaves of plants gathered by daylight be exis
of
tracted with alcohol to

tion
is
remove the chlorophyll, a distinct blue colorathem in iodine solution. This reaction dipping upon produced
not obtained, however, with leaves which are plucked bewhich proves that light energy is necessary for the formation of starch in the leaf and that the starch which is thus formed is
for starch is
fore daylight;
afterwards hydrolyzed into sugar.
which
The sugar, Transportation and Metabolism of Sugars in Plants. is produced in the leaf is afterwards transported to various
parts of the plant, where it is either transformed into cellulose, hemicellulose, and other substances of the mechanical tissue, or else stored
up
as reserve material in the
form of sucrose, starch,
inulin,
and other
carbohydrates.
The
plants. which are
intensity of assimilation has been measured for many different The results are usually expressed in grams of starch or sugar
formed per square meter of leaf surface in an hour. The determinations show differences for different plants and for different conditions of temperature and sunlight, the results varying from traces up to two grams or more of carbohydrates per square meter of leaf area per
hour.
in fact as
Measurements of sunshine, temperature, and leaf area are used a means of forecasting the probable production of sugar by a
beet crop.
CHAPTER XIX
THE MONOSACCHARIDES
DlOSES
CH
2
Glycolose.
Glycolaldehyde.
CH OH CHO
2
Glycolose has not been found as yet free in nature. It has been pre* pared synthetically by oxidation of its alcohol glycol with nitric acid
and by
electrolysis f
from glyceric
acid.
CH OH
2
HOH COOH
Glyceric acid
CH OH
2
CHO
Glycolose
+ C0 + H
2
Glycolose
is
also obtained
by the condensation
other ways.
of
two molecules
of
formaldehyde and in
95
Properties. to 97 C.,
It
is
many
is
Glycolose crystallizes in colorless plates, melting at easily soluble in water and alcohol and has a sweet
glycollic acid,
taste.
optically inactive
oxidation
first
monobasic
and unfermentable. It yields upon and then dibasic oxalic acid.
Tests. Glycolose gives all the ordinary sugar reactions. a-Naphthol and sulphuric acid give a bluish violet coloration { with total absorption of the red and violet parts of the spectrum and a band between the D and E lines. It forms a number of osazones of which the p-nitrophenyl-
osazone
is
especially characteristic the compound is very insoluble in the ordinary solvents but can be crystallized from pyridine; its melting
is
;
point
311
C.
METHYLDIOSES
CH C H
S 2
Methylglycolose.
Lactic aldehyde.
CH CHOH CHO
3
Fischer and Tafel., Ber., 20, 1091 22, 96. t Neuberg, Biochem. Zeitschr., 7, 527. t Neuberg, Z. Ver. Deut. Zuckerind., 61, 271.
;
635
536
SUGAR ANALYSIS
Lange* by saponifying
This, the simplest of methyl sugars, was prepared by Wohl and its acetal derivative with dilute sulphuric acid.
CH
CHOH
HC
The sugar
form (C 3 H6
/ OC 2 H 6
+H
\OC H
2
CH CHOH + CHO
3
C H OH
2 5
Lactic diethylacetal
Lactic aldehyde
Alcohol
as thus
split off is
obtained in a polymerized bimolecular
2 )2
consisting of large needles melting at 101
heating this polymerized obtained.

Properties
C.; upon compound the simple monomolecular sugar is
with slightly rancid odor.
Methylglycolose consists of a colorless liquid It is colored brown by alkalies, reduces and gives the other reactions of a simple reducing Fehling's solution, Its phenylhydrazone forms colorless leaflets melting at 92 C.; sugar.
Tests.
its
and
nitrophenylhydrazone consists of bright yellow prisms melting at Its osazone is identical with that of acetol and methyl129 C.
glyoxal.
its
While methylglycolose has not thus far been found free in nature monobasic acid derivative lactic acid, CH 3 CHOHCOOH, is very
widely distributed.
Acetol.
Acetylcarbinol.
CH
i-o
CH OH
2
This, the simplest of ketose sugars, can be prepared in a number It is formed by oxidizing a-propylene glycol with bromine of ways. or by the action of Bacterium xylinum.] water,
CH CHOH CH OH
3
2
CH
+ O
CO
-f
2
HO
2
CH OH
Acetol
o-Propylene glycol
Acetol
is also formed in large amounts by distilling glucose with very concentrated potassium hydroxide solution. Acetol consists of a colorless, sweet-smelling liquid Properties. with a nutty flavor, which boils in vacuum at 105 C. and in air at *
Ber., 41, 3612.
t Kling,
Compt.
rend., 128, 244; 129, 219, 1252; 133, 231.
THE MONOSACCHARIDES
147 C. with decomposition. It is easily soluble in water, alcohol ether and reduces Fehling's solution strongly in the cold. Acetol gives the oxime, hydrazone, osazone Reactions.
other reactions
537
and
and
common
Ci 5 Hi 6
4,
is
formed by
to reducing sugars. heating acetol with
The phenylosazone,
phenylhydrazine and
consists of yellow needles melting between 145 and 148 C.; the compound is identical with the osazone of lactic aldehyde and methyl-
glyoxal
(CH CO COH).
3
Acetol-phenylosazone
is
also
formed
acetol being
upon heating glucose with phenylhydrazine in alkaline solution, the first formed as a decomposition product of the glucose and then reacting with the phenylhydrazine. DlMETHYLDIOSES
(CH3 C 2 H
)2
Dimethylglycolose.
Dimethylketol.
Acetylmethylcarbinol.
CH CHOH
3
Ao
CH
Occurrence.
Dimethylglycolose
is
formed in small amounts

It
is
in
many
aerobic fermentations of sugars.
common
constituent of
cider vinegar f
and
Synthesis.
a frequent by-product in the acetic fermentation. Dimethylglycolose was prepared synthetically by Pechis
mannj by reducing
diacetyl with zinc
and sulphuric
3
acid.
CH
CO
CO
CH CHOH
~CO
3
CH
CH
Diacetyl
Dimethylglycolose
Properties. Dimethylglycolose is a colorless liquid boiling at 141 to 142 C., and is easily soluble in water and alcohol. Similar to other of the diose it is sugars group easily polymerized.
Dimethylglycolose reduces Fehling's solution even in the by means of its yellow finely crystalline which is very insoluble in water, alcohol, and Ci Hi 6 8 N4, phenylosazone, the also is ether; distinguished by its high melting point, compound
Tests.
cold.
It is best recognized
*
t
Pinkus, Ber., 31, 31.
Browne, J. Am. Chem. Soc., 26, 31. J Ber., 21, 2754; 22, 2214; 23, 2421.
538
SUGAR ANALYSIS
245 C., which seems to be the highest of any phenylosazone thus far
prepared.
TRIOSES
C3 H
ALDOTRIOSES
d, 1-Glycerose.
Glyceric aldehyde.
CH OH
2
HOH
CHO
Glycerose has not been found free in nature, but has been prepared * synthetically by oxidation of its alcohol glycerol, by action of water upon acrolein dibromide, and in other ways.
form of
d,l-Glycerose crystallizes from methyl alcohol in the Properties. The compound shows a colorless needles melting at 138 C.
It is optically inactive. great tendency to polymerize. tation of glycerose sirup by yeast, observed by Fischer
The fermenand
Tafel,
is
probably due to the formation of a fermentable condensation product. Pure glycerose according to Wohl f and Emmerling { is not fermentable. Tests. Glycerose reduces Fehling's solution and exhibits all the other reactions common to sugars. Heating with concentrated hydrochloric acid and a little orcin produces a bluish green color which soon
separates as a flocculent precipitate; solution of the latter in gives a characteristic absorption band between the C and
II
amyl alcohol
lines of the
spectrum. Phloroglucin in presence of a little sulphuric acid gives a flocculent precipitate with dilute glycerose solutions upon warming.
d, 1-Glycerose gives glycerol
first
upon reduction, and upon oxidation
monobasic d, 1-glyceric acid and then dibasic tartronic acid. The sugar has not been resolved as yet into d- and 1-glycerose; although d, 1-glyceric acid has been separated by Frankland and Frew If by fermenting calcium d, 1-glycerate with Bacillus ethaceticus which attacks
only the 1-component.
KETOTRIOSES
Dioxyacetone.
CH OH
2
i-o i H OH
2
Fischer and Tafel, Ber., 20, 3384.

||
t Ber., 31, 1796, 2394. * Ber., 32, 544.
Neuberg, Z. Ver. Deut. Zuckerind., 61, 271, Wohl and Neuberg, Ber., 33, 3095.
J.
Chem.
Soc., 69, 96;
63, 296.
THE MONOSACCHARIDES
Dioxyacetone
is
539
formed as a by-product in a number of different been prepared synthetically in several ways, but the best method is that of Bertrand* which consists in fermenting
fermentations.
It has
a 5 or 6 per cent glycerol solution with Bacterium xylinum. When the reducing power of the solution has reached its maximum, fermentation
interrupted; the solution is evaporated in vacuum, the sirup extracted with 5 to 6 parts alcohol and 2 parts ether, and the dioxyacetone crystallized from the alcohol-ether extract.
is
Dioxyacetone is a white crystalline compound soluble water and boiling alcohol. It has a sweet taste and melts between 68 and 75 C. under polymerization. Its concentrated water solutions also polymerize readily yielding a crystalline compound of melting point 155 C. It is optically inactive and not fermented by
Properties.
in cold
yeast.
Dioxyacetone reduces Fehling's solution even in the cold. ketoses it gives the characteristic reaction with resorcinf and an osazone with methylphenylhydrazine. This osazonef has the formula Ci 7 H 2 oN 4 O and melts at 127 to 130 C. Distillation of dioxyacetone with 20 per cent sulphuric acid gives methylglyoxal, CH3 COCHO. Reduction with sodium amalgam gives glycerol quantitatively. Dioxyacetone does not give the reaction with phloroglucin
Tests.
Similar to
all
||
characteristic of the isomeric glycerose.
METHYLTRIOSES
CH C H
3 3
Methylglycerose.
CH HOH HOH
3
HO
This compound has been prepared synthetically by Wohllf and Frank from crotonaldehyde. It forms a colorless sirup easily soluble in water and alcohol and reduces Fehling's solution about half as strong
as glucose.
*
Compt.
rend., 126, 842, 984.

61, 271.
t
j
Neuberg, Z. Ver. Deut. Zuckerind., Neuberg, Ber., 36, 964.

Pinkus, Ber., 31, 31. Piloty, Ber., 30, 1656, 3161.
Ber., 35, 1904.
II
540
SUGAR ANALYSIS
TRIMETHYLTRIOSES
(CH3 ) 3 C3H3
Trimethyltriose.
CH CH
3
\ /
COH
CHOH
i-o CH
3
This compound, which belongs to the ketoses, has been made * from mesityl oxide. It consists synthetically by Harries and Pappos
of a bright yellow sirup of caramel-like odor, easily soluble in water, alcohol and ether.
TETROSES
C 4H
d-Erythrose.
ALDOTETROSES
CH OH
2
HOCH HOCH CHO

Wohl t through decomposition
solution.
This sugar has been prepared synthetically from d-arabonic acid by of the nitrile with ammoniacal silver
Rufff has also prepared the sugar by oxidation of calcium d-arabonate with hydrogen peroxide in presence of ferric acetate. In this reaction the COOH group of the acid is split off with evolution
of
C0
2.
CH OH
2
CH OH
2
HOCH HOCH HCOH COOH

d-Arabonic acid
HOCH
O = HOCH
+ CO + H O
2 2
CHO
d-Erythrose
is
The
configuration of d-erythrose
established
by means
of these
reactions.
Properties. d-Erythrose consists of a colorless sirup which solidito a white mass when dried over phosphorus pentoxide. It is
*
fies
Ber., 34, 2979.
f Ber., 26, 743.
J Ber., 32,
3672.
THE MONOSACCHARIDES
easily soluble in
541
optically active
water and alcohol.

14.5
The sugar
is
and
(+ aqueous solution). not fermented by yeast. Tests. d-Erythrose reduces Fehling's solution and gives all other reactions common to reducing sugars. Reduction with sodium amalgam
exhibits mutarotation; [O\D
1.0 in fresh
d-Erythrose
is
gives optically inactive mesoerythrite, which is widely distributed in nature in different algae and lichens. Oxidation of d-erythrose produces first monobasic d-erythronic acid and then dibasic mesotartaric
acid.
1-Erythrose.
CH OH HCOH HCOH
2
This sugar has been prepared synthetically from 1-arabonic acid according to the methods of Wohl* and Rufff described under d-erythrose. Neuberg J has also prepared the sugar from 1-arabonic acid by
his
method
of electrolysis.
1-Erythrose consists of a colorless sweet sirup which Properties. has not as yet been obtained crystalline. The sugar is dextrorotatory,
found [a] D = + 21.5 constant and in fresh solution +2.4. The differences noted are probably due to the fact that the sugar has not yet been isolated in the pure condition. 1-Erythrose is not fermentable.
[O\D
= + 32.7
(Wohl)
Ruff and Meusser
Tests.
1-Erythrose gives
all
the ordinary reactions of reducing
Reduction with sodium amalgam gives inactive mesoerythrite the same as d-erythrose; oxidation produces first monobasic 1-erythronic acid and then dibasic mesotartaric acid.
sugars.
Racemic erythrose d, 1-Erythrose. natural mesoerythrite.
is
formed by the oxidation
||
of
CH OH HCOH HCOH CH OH
2
|
CHO
+0
HCOH HCOH CH OH
|
+|
J
CH OH HCOH + 2H HCOH CHO

2
Mesoerythrite
d,l-Erythrose
Ber., 32, 3666.
f Ber., 34, 1366.

II
Biochem.
Zeitschr., 7, 527.
Ber., 34, 1366.
Fischer and Tafel, Ber., 20, 1090.
542
SUGAR ANALYSIS
first
The sugar is of course inactive. Oxidation produces erythronic acid and then mesotartaric acid.
1-Threose.
d,
CH OH HOCH HCOH
2
This tetrose sugar has been formed synthetically by oxidation * of calcium 1-xylonate with hydrogen peroxide and ferric acetate.
CH OH HOCH HOCH HCOH + O = + CO + H O HCOH HOCH CHO COOH

2 2
2
|
CH OH
2
l-Xylonic acid
1-Threose
The
Properties.
dition, all
Tests.
configuration of 1-threose is established by this reaction. 1-Threose has only been obtained in a sirupy con-
attempts to promote crystallization having
failed.
+4.25).
1-Threose upon reduction gives 1-erythrite ([]z> in water = Oxidation gives first 1-threonic acid and then 1-tartaric acid.
d-Threose. The optical antipode of 1-threose has not as yet been prepared. Its alcohol d-erythrite, however, has been obtained = 4.40) by reduction of d-erythrulose. ([O\D in water
KETOTETROSES
d-Erythrulose.
CH OH
2
HCOH
CH OH
2
by means
This sugar is best prepared by oxidation of natural mesoerythrite of Bacterium xylinum according to Bertrand's f method. Properties. d-Erythrulose has been obtained only as a sirup it is
;
very soluble in water and alcohol, and

increasing after solution.
Tests.
*
is
dextrorotatory, the rotation
The sugar is unfermentable. d-Erythrulose gives the ordinary ketose reactions, produc
t
Ruff and Kohn, Ber., 34, 1370.
Compt.
rend., 130, 1330.
THE MONOSACCHARIDES
543
ing a coloration with resorcin and hydrochloric acid and resisting oxidaReduction with sodium amalgam gives both tion with bromine water.
meso- and d-erythrite.
CH OH CH OH HCOH HCOH HCOH 21 +2H = + C=O HCOH HOCH CH OH CH OH CH OH

2
CH OH
d-ErythruIose
Mesoerythrite
d-Erythrite
This property of yielding two different alcohols upon reduction is a characteristic of the ketose sugars. d, 1-Erythrulose is formed according to Neuberg* during the oxidation of mesoerythrite by hydrogen peroxide in presence of ferrous sul-
phate (Fenton'sf synthesis). The sugar has been obtained only as a sirup and has not been solved as yet into its d- and 1- components.
re-
METHYLTETROSES
CH C H O
3 4
7
Methyltetrose.
CH
CHOH HCOH HOin

CH(
This sugar has been prepared synthetically by Fischer | from rhamnonic-acid nitrile
by Wohl's method. has not been obtained in a pure crysMethyltetrose Properties. talline form, but only as a yellowish sweet sirup easily soluble in water
and alcohol with levorotation,
Tests.
[a]o
5M
(in water).
sugar.
Methyltetrose gives the ordinary reactions of an aldose Reduction with sodium amalgam gives methylerythrite. Ox-
idation with bromine gives methyltetronic acid, whose lactone gives [a] D = - 47.5. Oxidation with nitric acid splits off the 3 group with for-
CH
mation of d-tartaric
acid.
DlMETHYLTETROSES
(CH 3 ) 2 C4H 6 O 4
Digitoxose.
tetrose,
C Hi2
6
4,
This sugar which has the composition of a dimethyl has been obtained by Kiliani by hydrolysis of digit Ber., 29, 1377.
Ber., 35, 2627.
f J,
Chem.
Soc., 71,
375 (1897).
Ber., 31, 2454;
34, 3561.
544
toxin,
SUGAR ANALYSIS
a glucoside found in different plants of the digitalis family. formation of digitoxose is supposed to proceed as follows:
The
C3 4
H On + H
54
= CaH04 + 2 C H
6
12
4.
Digitoxin
Digitoxigenin
Digitoxose
soluble
Digitoxose has been obtained as prismatic crystals melting at 101 C., in water and alcohol, and having a specific rotation of
Tests.
kb=+46.
Oxidation with silver oxide gives among other products Heated with concentrated sulphuric acid and 1 per cent ferrous sulphate, digitoxose solutions are colored, after 30 minutes, a deep blue, which changes in an hour or two to bluish green.
considerable acetic acid.
OXYMETHYLTETROSES
CH OH C H
2 4
Apiose
/3-Ox"ymethyltetrose.
CH OH
2
HOC-CH OH CHOH
2
CHO
This sugar, which has the same empirical formula C 5 Hio05 as a * pentose, has been found by Vongerichten as a constituent of the gluApiin upon treatcoside, apiin, which occurs in the parsley plant.
ment with strong
acids
is
2
hydrolyzed as follows:
C 26 H
80 14
+ 2H
=C
10
Apiin
Apiose
H + C 5H +C d-Glucose
6
12
10
5.
Apigenin.
Apiose has been obtained only as an optically inactive, unfermentable sirup.

Tests.
Apiose
is
distinguished from the pentose sugars
by not
giving furfural
upon heating with hydrochloric acid.
Reduction with
hydriodic acid and phosphorus gives iso valeric acid,
CH
HC-CH HCH
COOH,
which confirms the branched structure of the carbon chain assigned to

apiose.
*
Ann., 318, 121; 321, 71.
THE MONOSACCHARIDES
PENTOSES
ALDOPENTOSES
d-Arabinose.
545
CH OH HOCH HOCH OH
2
i HO
This sugar, which has been found in nature only as a constituent of d,l-arabinose in abnormal urines, has been prepared synthetically by WohPs* method from the nitrile of d-gluconic acid and by Ruff's f
method from the calcium

2
salt of d-gluconic acid. d-gluconic acid to d-arabinose proceeds as follows:
The
oxidation of
CH OH HOCH HOCH HOCH + O = HOCH + CO + H O HCOH >H HC01 HOCH CHO COOH
2
CH OH
d-Gluconic acid
d-Arabinose
is
The
reaction.
configuration of d-arabinose
established
by means
of this
Properties. melting at 160

alcohol.
d-Arabinose consists of beautiful prismatic needles C. and easily soluble in water, but insoluble in absolute
in
The sugar shows
aqueous solution
(c
9.45) [a] D
105
(constant); mutarotation is present; d-arabinose is not fermentable. Tests. d-Arabinose reduces Fehling's solution, yields furfural upon
distillation
teristic of
with hydrochloric acid and gives the other reactions characan aldopentose sugar. Reduction with sodium amalgam gives = + 7.7 (in saturated borax solution). d-arabite, C 5 Hi 2 O 5 for which [a]n Oxidation with bromine gives d-arabonic acid, whose lactone C 5 H 8 5
,
gives [d\D
=+
73.73.
glutaric acid, [O\D
=+
22.8.
Oxidation with strong nitric acid gives d-trioxyEspecially characteristic of d-arabinose is
the very difficultly soluble 1-menthylhydrazone J which separates in

*
Ber., 26, 730.

32, 550;
t Ber., 31, 1573; J
33, 1799;
36, 2360.
Neuberg, Ber., 36, 1194.
546
SUGAR ANALYSIS
C. and from which d-arabinose can
colorless crystals melting at 131
be isolated by decomposition with formaldehyde.
L-ARABINOSE.
CH OH HCOH
2
HC01 )H
HOCH CHO
Occurrence.
Ordinary or 1-arabinose has not been found free in
nature except as a constituent of d, 1-arabinose in abnormal urines; parent substances, from which 1-arabinose may be derived by hydrolyChief of these are, however, very widely distributed in nature. parent substances is the pentosan araban (C 5 8 O4) n which occurs as a constituent of many plant gums (cherry gum, peach gum, gum arabic, gum tragacanth, etc.), of the hemicellulose tissues of vegetable
sis,
cells
of
(sugar beet, maize stalks, elder pith, sugar cane, bran, etc.), and many plant mucilages (such as quince) and pectins. 1-Arabinose has
also
been found in several glucosides.
The Arabans. Araban itself (C 5 H 8 04) n occurs in nature not so much in the free condition as in a combined or associated form. The
chemistry of this group of substances
satisfactory classification
is
is
exceedingly complex and a
impossible. Among the arabans, or substances which yield 1-arabinose upon hydrolysis, are metaraban, glucoaraban, galactoaraban, arabinic acid, pectose, pectin, parapectin,
metapectin, parapectic and metapectic acids, and many other ill-defined substances. The early investigators in this field were hampered
by a lack
identical.
of satisfactory
methods and many

if
of the substances, to
they gave separate names, would,
purified,
which no doubt prove to be
A comparatively pure araban has been prepared by digesting sugarbeet pulp,* and other hemicelluloses,f with dilute alkalies. The clear filtrate is precipitated with weak acids in presence of alcohol. The
precipitate, after washing with strong alcohol, is purified by dissolving in water, and reprecipitating with alcohol. The product, after drying, consists of a white amorphous mass, soluble in water to a neutral solution,
([<*]D
does not reduce Fehling's solution and is strongly levorotatory 84 to given by different authorities varies from 123; these
* t
Ullik, Oest.
Ungar. Z. Zuckerind., 23, 268. Schulze, Z. physiol. Chem., 16, 386.
THE MONOSACCHARIDES
547
variations are probably due to differences in the purity of the product). Upon heating with 1 per cent sulphuric acid araban is quickly hydrolyzed to 1-arabinose.
(C 5 H 8
Araban
4 )n
+ nH
= nC
10
5.
1-Arabinose
found in the bran of rye, wheat and other cereal The bran, after removing the starch, is heated 3 hours with grains. The 1 per cent ammonia, and then filtered and washed with water. with sodium under dilute which residue is then cooked hydroxide pressure The latter is precipitated from the filtered dissolves the metaraban.
Metaraban*
is
solution
by means
of dilute hydrochloric acid
and
alcohol.
The
precipi-
tate, after washing with alcohol and drying, forms a white amorphous substance, which swells up in water and finally gives a mucilaginous,
slightly levorotatory solution.
Hydrolysis with acids gives 1-arabinose. Arabinic Acid"\ (arabin, metapectic acid) occurs in combination with potassium, calcium and magnesium as the principal constituent of gum
arabic and the
gum
of the cherry, peach,
plum and many other
trees.
It is also produced by the action of alkalies upon pectose and other Arabinic acid can be prepared by dissolving gum pectin substances.
arabic t in 10 parts of water, acidifying with acetic acid to break up mineral combinations and then dialyzing, or washing, in acetic-acid
The product salts and other impurities. with water and alcohol; it is precipitating purified by then dried over sulphuric acid at a low temperature, preferably in a
solution to
is
remove soluble
dissolving in
vacuum.
Arabinic acid can also be prepared, but in a less pure condition, by The latter, after extraction the action of alkalies upon beet pulp. with water and cold 85 per cent alcohol, is boiled in water until all
alcohol
is
expelled
and then cooked with an excess
of caustic lime.
The solution of lime arabinate is filtered and the lime precipitated by means of carbon dioxide, or oxalic acid. The solution is again filtered and the arabinic acid precipitated by adding an excess of strong alcohol. The crude acid is purified and dried as previously described.
Arabinic acid
being dried
it is
a white vitreous amorphous substance. Before easily soluble in water to an acid solution; but the dry
is
a product swells up with water to a mass of almost neutral reaction its lactone. acid into of the a conversion to attributed which is change
Ber., 23, 3110. prakt. chem., 62, 193 (1854), Scheibler, Ber., 1, 58; 6, 612. t O'Sullivan, J. Chem. Soc., 45, I, 41, Proc. Chem. Soc., 17, 156. Scheibler, Z. Ver. Deut. Zuckerind., 23, 288.
*
Steiger
and Schulze,
J.
Neubauer,
548
SUGAR ANALYSIS
80 to [a]o of arabinic acid of different origins varies from over Distillation with hydrochloric acid gives large amounts of
The
over +80.
furfural
and oxidation with
nitric acid considerable
mucic
acid.
Herz-
feld* obtained from a levorotatory arabinic acid 15.3 per cent furfural and 11.5 per cent mucic acid and from a dextrorotatory arabinic acid
5.9 per cent furfural
and 41.7 per cent mucic acid. It is thus seen that a galactoaraban of varying composition. Hydrolysis of both levorotatory and dextrorotatory arabinic acid gives a dextroarabinic acid
is
rotatory mixture of 1-arabinose and d-galactose. Neubauer assigned 'Sullivan has given the the formula (Ci 2 2 20ii) n to arabinic acid;
formula
iHi4 2 O 7 4.
Such
differ-
ences necessarily follow from the variable character of the substance.

arabinic
Metarabin is obtained by heating acid to slightly above 100 C., at which temperature water
is given off. It is insoluble in water, yielding only a swollen gelatinous
mass.
as (Ci 2
The formula has been given
20 Oio)n.
Other mixed arabans, as arabogalactan and the pectin substances, are described under d-galactose.
Preparation of 1-Arabinose. 1-Arabinose can be prepared by the hydrolysis of araban, metaraban or

arabinic acid; it is more convenient, however, to prepare the sugar by the direct hydrolysis of certain
gums.
Cherry
gum
is
one of the
Tollens's apparatus for hyFig. 190. drolyzing plant and animal substances.
purest sources of supply, and as the preparation of arabinose from this
substance
drolytic
is
typical of other hy-
processes,
the
following
be described in fuller detail. Gum. Treat 1000 gms. of pulverized cherry Hydrolysis of Cherry in a c.c. of water and 280 gms. of conwith 7000 gum large porcelain pot centrated sulphuric acid, thus making a mixture of about 4 per cent
of Tollensf will
acid.
*
method
The pot
Z. Ver.
is
immersed
in a boiling-water bath, as
shown
in Fig.
"
t
Handbuch
Deut. Zuckerind., 41, 667. " der Biochemischen Arbeitsmethoden (1902), Vol.
II, 65.
THE MONOSACCHARIDES
549
The pot is 190, and the mixture stirred until the gum has dissolved. then covered and the heating continued for 5 hours. The liquid, which smells strongly of furfural, is then poured into a large evaporathot neutralized with an excess (300 to 320 gms.) of precipitated calcium carbonate, which must be free from
ing dish,
still
and while
Fig. 191.
Tollens's apparatus for evaporating sugar solutions under reduced pressure.
magnesium carbonate (the magnesium sulphate which is formed interfering with the crystallization of the sugar). The liquid after cooling is filtered through a heavy linen bag, and the precipitate of gypsum, etc., squeezed out in a press (Fig. 131) to remove as much as possible of the
liquid.
contains in addition to arabinose
The hydrolyzed solution reduces Fehling's solution strongly, and more or less galactose and glucose. In
order to remove the latter, the solution is poured into a large bottle and fermented in a warm place with a little pure pressed yeast. When
is complete (3 to 4 days at most), the solution is filtered and evaporated under diminished pressure to a sirup. In conducting the evaporaEvaporation Under Reduced Pressure. tion of sugar solutions a small laboratory vacuum pan may be used to advantage. In place of such a pan the arrangement of Tollens shown in Fig. 191 may be used to equal advantage. The liquid is placed in
fermentation
550
SUGAR ANALYSIS
the large balloon flask F of heavy glass, which rests in an inclined The flask is closed with a twoposition upon the hot-water bath W. receives which hole rubber stopper, through one opening the tube t; the latter, drawn out to a fine point, reaches nearly to the bottom of the flask and is fitted at the outer end with a rubber tube and pinch cock c'. The flask is connected by the bent tube c to a vertical con-
which fits into a large Woulf bottle B. The latter is connected one side with a closed outlet tube G and upon the other side with a upon S to which the suction pump is attached. bottle safety In making an evaporation the pump is started and a gentle current of air drawn through the liquid while the bath is being heated. By diminishing the air, the pressure is reduced so that the solution soon begins to boil. The current of air is always maintained slightly so as
denser,
to keep the liquid in motion and prevent bumping. to empty the receiver the pinch cock c' is opened
When
it is
desired
and the
distillate
siphoned
off at
the outlet G.
When the liquid in the flask has been conPurification of Sirups. centrated to a sirup, the latter is poured out and a fresh quantity of The concentrated sirups are then united and solution evaporated.
shaken with 4 to 5 volumes of hot 96 per cent alcohol. After the deposit of gums and mineral matter has settled, the alcoholic solution is If filtered and evaporated under reduced pressure to a second sirup. the latter be very dark in color, it may be further purified by shaking out again with warm alcohol to which a little ether may be added to An excess of ether must be avoided increase the precipitation of gum. The final sirup, which should be as it precipitates part of the sugar.
light colored,
is
set aside in a cool place.
Crystallization.
The
is
crystallization of arabinose
it
from sirups pre-
may be hastened by primgum usually rapid; minute of with a the sugar from a stock preparation. ing crystal sirup When crystallization is complete, the crystals of sugar are sucked off
pared from cherry
upon a
filter,
washed with a
little
alcohol
and
ether,
and
air dried.
If
the sugar is not perfectly white, it may be purified by recrystallizing from hot alcohol after filtering through bone black. The yield of 1-arabinose from cherry gum by the above process is about 20 per cent.
1- Arabinose crystallizes in beautiful prismatic needles Properties. melting at 160 C., easily soluble in water but insoluble in absolute alcohol and ether. The sugar shows strong mutarotation [O\D = 104.5
;
(constant in aqueous solution).

1- Arabinose is
able to set
not fermented by yeast; many bacteria, however, are up destructive changes with formation of lactic, acetic,
THE MONOSACCHARIDES
formic, succinic, oxalic and other acids. the sugar to 1-arabonic acid.
551
Bacterium xylinum oxidizes
Tests. 1-Arabinose gives all the general reactions described for reducing sugars and the furfural, color and other special reactions de-
The best method for detecting 1-arabinose in presence of other sugars (as in hydrolyzed plant materials) is by means of the hydrazone reaction with different substituted hydrazines,
scribed for pentoses.
such
as
bromophenylhydrazine,*
methylphenylhydrazine,|
benzyl-
The latter reagent is phenylhydrazine % and diphenylhydrazine. considered to be the best and produces in alcoholic solution in the cold, even with small amounts of 1-arabinose, a difficultly soluble hydrazone, Ci7H 2 oN 2 04, consisting of white needles and melting at 204 to 205 C.
||
The hydrazones
maldehyde
of 1-arabinose yield,
upon decomposition with
for-
(p. 348), the free sugar which may then be crystallized further identified by determining its specific rotation.
and
for
Sodium amalgam reduces

which
[O\D
1-arabinose
to
1-arabite
C Hi O
5 2
5,
5.31f (in saturated
ammonium
molybdate).
borax solution), and 42 ** (in acid Oxidation of 1-arabinose with bromine gives
1-arabonic acid,
acid, [a] D
whose
crystalline lactone
CHO
5
gives [O\D
73.9.
Oxidation of the sugar with strong nitric acid gives 1-trioxyglutaric
=-
22.7.
This sugar, which is a racemic mixture of d- and been found as a constituent ft f abnormal urines. The sugar may also be prepared by dissolving equal parts of d- and 1-arabinose in hot alcohol and allowing the solution to crystallize. d,l-Arabinose forms colorless prismatic crystals of Properties. higher melting point (164 C.) and lower solubility than either of its components. The sugar is optically inactive and is not fermented by
d, 1-Arabinose.
1-arabinose, has
yeast.
Tests.
d, 1-arabite;
Reduction of
d, 1-arabinose
with sodium amalgam gives
oxidation with bromine gives d, 1-arabonic acid and with nitric acid d, 1-trioxyglutaric acid. Neuberg J J has resolved the sugar into its components by means of 1-menthylhydrazine, which forms a very
difficultly soluble
hydrazone with d-arabinose, but not with 1-arabinose
(see
*
pages 362 and 545).

Fischer, Ber., 24, 4214; 27, 2490. Ruff and Ollendorff, Ber., 32, 3234. Ruff and Ollendorff, Ber., 32, 3234. Neuberg, Ber., 33, 2254; 37, 4616.
H Fischer,
**
Ber., 24, 1836.
t
t
Gernez, Compt. rend., 112, 1360. ft Neuberg, Ber., 33, 2243.

tt Ber., 36, 1194.
II
Muther and
Tollens, Ber., 37, 312.
552
d-Xylose.
SUGAR ANALYSIS
CH OH HCOH HOCH HCOH CHO
2
or combined form.
This sugar has not thus far been found in nature either in the free It has been made synthetically by Fischer and
Ruff* from d-gulonic acid by oxidation of the calcium salt with hydrogen peroxide in presence of ferric acetate (Ruff's method).
CH OH
2
1
+ C0 + H O
2 2
THE MONOSACCHARIDES
from which 1-xylose
553
may be
derived by hydrolysis, are, however,
among
the most widely distributed substances in the vegetable world. Chief of these parent substances is the pentosan xylan (CsHgOJn or wood
gum, which occurs as a constituent of the incrusting or hemicellular materials which are found in nearly all vegetable cells. Xylan with a little araban makes up from 25 to 30 per cent of the dry matter of cereal straws and grasses, about 15 to 25 per cent of the dry matter of the wood of deciduous trees and from 5 to 15 per cent of the dry matter of the wood of coniferous trees. It is also found in large in bran of seeds and quantities bark, roots, grains, mosses, fungi and associated with araban as a constituent of many vegetable gums. Xylan is, next to cellulose, the most abundant of plant constituents.
One of the best sources for preparing Preparation of Xylan. is beech-wood xylan sawdust, although maize stalks, straw and other materials plant may be used. According to the method of Wheeler and Tollens* 1 kg. of fine beech-wood sawdust is first treated in the cold for 24 hours with 1 to 2 per cent ammonia to dissolve albuminoids,
the ammoniacal solution is then pressed out and the process repeated for a second or third time. The material after washing with water is then treated in a warm place with 5 per cent sodium hydroxide
etc.
;
solution, the latter being
added
in sufficient quantity to
is
form a thick
mush.
After 24 hours the extract
pressed out and the digestion re-
extracts are
peated for another 24 hours using less sodium hydroxide solution. The mixed and allowed to stand in a flask for deposition of sus-
pended impurities. The clear brown colored solution is then siphoned and mixed with an equal volume of 96 per cent alcohol which preThe latter is filtered cipitates the xylan as a sodium-gum compound. off upon cloth, washed with alcohol till the washings are colorless and
off
then treated in presence of alcohol with hydrochloric or acetic acid until the reaction is slightly acid. The free xylan, which is thus liberated, is washed first with alcohol upon cloth, or parchmentized paper, using suction, until all acid is removed, then washed with a little ether, and finally dried over concentrated sulphuric acid. The product thus prepared consists of a grayish white amorphous powder, which is almost insoluble in water. 70 In alkaline solution it is levorotatory, [O\D = to 90 according to the purity and origin of the gum. Xylan upon heating with dilute hydrochloric or sulphuric acid is quickly hydrolyzed
to 1-xylose.
(C 5 H 8
4) n
+nH
= n C Hi
5
5.
Xylan
l-Xylose
* Z. Ver. Deut. Zuckerind., 39, 848, 863.
554
SUGAR ANALYSIS
in the
The Xylo-proteids. 1-Xylose has also been found widely distributed animal world as a constituent of many nucleo-proteids. The
latter are
by hydrolysis
complex compounds of variable composition and are resolved into a mixture of substances, among which the nitrog-
enous bases (adenine, guanine, xanthine and hypoxanthine), phosphoric acid and various sugars have been identified. 1-Xylose seems to be
the most abundant of the sugars entering into the composition of the nucleo-proteids although other pentose and hexose sugars have been identified. The amount of pentose sugar in different organs has
been found by Grund
to be 0.021 per cent in muscle, 0.090 per cent
in the brain, 0.081 per cent in the spleen, 0.084 per cent in the kidney, 0.110 per cent in the liver, and 0.447 per cent in the pancreas; it is
especially in the pancreas that the occurrence of 1-xylose in the animal body is localized. The origin of 1-xylose in the animal organism is not
absolutely known although Neuberg and Salkowski f regard d-glucuronic acid as the parent substance from which it is derived. The nucleo-proteids are also widely distributed in the vegetable and give the same products upon hydrolysis.
kingdom
Preparation of 1-Xylose.
1-Xylose
may be
prepared by hydrolysis
of xylan obtained as described above or by direct hydrolysis of plant materials. Xylan may be hydrolyzed with sulphuric acid in the same
way
to Councler's { method.
gum or with hydrochloric acid according For the latter process 15 gms. of xylan are heated on the water bath with 200 c.c. water and 70 c.c. hydrochloric acid (1.19 sp. gr.) for 3 hours. The solution is then treated with pure lead carbonate, until Congo-red test paper shows no free acid, and filtered. The filtrate is evaporated to a thin sirup in presence of a little lead carbonate and then treated with strong alcohol to precipitate gums, lead chloride and other impurities. The alcohol solution is treated with hydrogen sulphide to precipitate any remaining lead, filtered and evaporated to a sirup in presence of a little calcium caras described for cherry
bonate.
cool place
crystals
The bright straw-colored sirup thus obtained is set aside in when crystallization of xylose will proceed rapidly. The are purified by recrystallizing from alcohol using a little animal
charcoal.
1-Xylose can also be prepared directly from wheat straw, maize The material is stalks, etc., by the process of Schulze and Tollens.
purified by digesting with 2 per cent ammonia and water (as described under preparation of xylan), and then, after pressing as dry as
first
*
Z. physiol. Chem., 36, 111.
Chem.
Ztg. (1892), 1719.
t Z. physiol.
Chem.,
36, 261;
37, 464.
Ann., 271, 40.
THE MONOSACCHARIDES
555
possible, heated on the water bath 4 hours with 2 to 3 per cent sulphuric acid. The acid extract is pressed out, neutralized with pow-
dered calcium carbonate, filtered and evaporated (as described under preparation of 1-arabinose) to a sirup. The latter, after shaking out
several times with hot 96 per cent alcohol to precipitate gums, yields upon evaporation 3 to 5 per cent of the weight of straw in crystallized
xylose.
1-Xylose crystallizes in white needles of Properties of 1-Xylose. a sweetish taste, which are easily soluble in water and hot alcohol, but The sugar melts at about 145 C., although differinsoluble in ether.
ent authorities vary 10 C. above or below this figure owing no doubt
to variations in method.
1-Xylose shows stronger mutarotation than other 5 minutes after solution = 85.68 (Wheeler and v any sugar; [a]n Tollens *) [a]o constant = 18.5. The sugar is not fermented by
yeast; many bacteria and moulds, however, are able to produce destructive changes with formation of lactic, succinic, acetic and 1-xylonic
acids.
1-Xylose gives all the general reactions described for reducing sugars and the furfural and other special reactions described for the pentoses. One of the best methods for detecting xylose in
Tests.
presence of other sugars and cadmium carbonate.
Bertrand's f reaction by means of bromine oxidizes the xylose to xylonic acid according to the following reaction:
is
The bromine
CH OH
2
CH OH
2
(CHOH) 3
H2 O + Br2 = (CHOH) 3
COOH
Xylonic acid
HBr
CHO
Xylose
Hydrobromic acid
The
xylonic and hydrobromic acid react with the cadmium carbonate forming cadmium xylonate and bromide, the solution of which on evaporation deposits characteristic boat-shaped crystals of the double bromide and xylonate of cadmium (C 5 9 O 6 )2Cd CdBr2 2 H 2 O. The
salt
can be purified by recrystallizing and should show upon analysis
29.86 per cent Cd and 21.32 per cent Br. Bertrand's reaction, according to Tollens and Widtsoe,t is carried out as follows. For each 0.2 gm. sugar or double the amount of sirup
to be tested, 1
c.c.
of water, 0.25
gm. bromine
(7 to 8 drops)
and
0.5
gm.
cadmium carbonate are mixed together in a test tube with gentle warming
*
Ber., 22, 1046.

[3],*
t Bull. soc. chim., J Ber., 33, 132.
6,
546.
556
SUGAR ANALYSIS
and then, after corking loosely, set aside for 24 hours. The solution is then evaporated in a dish almost to dryness, taken up with a little water, If xylose is present filtered and again evaporated almost to dryness. addition of a little alcohol will soon cause crystals of the double cadmium salt to deposit. Presence of impurities may delay the crystallization somewhat. Too much bromine must be avoided in making the test,
and an excess
of
cadmium carbonate must always be
present.
The
first
crop of crystals frequently appear amorphous, but the characteristic boat-shaped needles are always obtained upon recrystallizing. A second method which has been employed for the detection of
1-xylose in
impure mixtures
is
which separates
in crystalline
by means form upon
of the diformal *
compound,
boiling xylose solutions with
^paraformaldehyde (trioxy methyl ene). 1-Xylose-diformal has the formula C 5 H 6 05(CH2)2 and consists of white crystals melting at 56 C.; it can be sublimed without decomposition and shows in methyl alcohol
H>=+25.7.
1-Xylose upon reduction with sodium amalgam is converted into the Oxidation with bromine gives optically inactive pentite alcohol, xylite. nitric acid inactive xylotrioxyglutaric acid. 1-xylonic acid and with
1-Xylose has been synthetized from 1-gulonic acid by Fischer and Ruff f employing the same method described for d-xylose.
Racemic xylose has been prepared by Fischer and Rufft by crystallizing a mixture of equal parts of d- and 1-xylose out of 96 per cent alcohol. The sugar consists of small prismatic
d, 1-Xylose.
Its phenylosazone melts at to 131 C. crystals melting at 129 210 to 215 C., whereas the phenylosazone of 1-xylose melts at only
160 C.
d, 1-Xylose
is
also
formed by the oxidation of inactive xylite by
means
of bromine.
CH OH HOCH 2 HCOH + HOCH CH OH

2
2
CH OH HOCH HOCH = HCOH + HCOH + 2 H HOCH HOCH CH OH CHO

CHO
2
2
Xylite
d-Xylose
1-Xylose
Lobry de Bruyn and van Ekenstein, Rec.
trav. Pays- Has, 22, 159.
t Ber., 33, 2142.
t Ber., 33, 2145.
THE MONOSACCHARIDES
Xylose has not been resolved as yet into components.
d-1
its
557
optically active
d-Lyxose.
CH OH HOCH HCOH C
2
HC<
d-Lyxose has not been identified with certainty in any natural product, although Haiser and Wenzel* believe to have obtained it in the hydrolysis of inosinic acid (a nucleic acid found in meat extract). The
sugar has been
made
synthetically in a variety of ways;
by reduction
of d-lyxonic lactone,
(Wohl'sf method) and by oxidation of calcium d-galactonate by hydrogen peroxide in presence of ferric acetate (Ruffst method).
by degradation
of d-galactonic nitrile
CH OH
2
C0 + H
2
558
SUGAR ANALYSIS
1-Lyxose and d, 1-lyxose have not as yet been prepared.
d-Ribose.
CH OH
2
HOCH HOCH HOCH CHO

* regarded by Levene and Jacobs as a constituent of many Inosinic acid nucleic acids in the animal and vegetable kingdom. the has authorities these to configuration. according
This sugar
is
N-C-N
H H H H O = P-O-CH -C-C-C-C - N-C-CH
/
2
OH
CH
OH
OC-NH
Phosphoric
acid radical
d-Ribose
radical
Hypoxanthine
radical
Hydrolysis of the above at nearly neutral point produces free phosphoric acid and the d-ribose-hypoxanthine base; the latter upon further hydrolysis is decomposed into free d-ribose and free hypoxanthine. Levene and Jacobs f have also obtained d-ribose from guanylic
acid and yeast nucleic acid. d-Ribose as prepared by Levene Properties. colorless crystals melting at 85 C. and giving []
and Jacobs forms
=-19.25.
The
bromophenylhydrazone forms white needles melting at 170 C.

1-Ribose.
CH OH
2
HCOH HCOH HCOH CHO

This sugar has not been found as yet in any natural product; it has been prepared J synthetically by reducing the lactone of 1-ribonic
* J.
Am. Chem.
Soc., 32,
231;
Ber., 42,
1198.
This view of Levene and
Jacobs
j
contested, however, t Ber., 42, 2474.
is
by Neuberg,
Ber., 42, 2806.
Fischer and Piloty, Ber., 24, 4214.
THE MONOSACCHARIDES
acid which can be prepared from 1-arabonic acid pyridine (p. 775). From the sirupy mixture obtained
1-ribose
559
by heating with by this reduction
can be precipitated as the phenylhydrazone or bromophenylhydrazone. By decomposition of the latter with benzaldehyde van Ekenstein and Blanksma* were able to isolate the sugar in the pure
crystalline form.
1-Ribose consists of white needles melting at 87 C., Properties. in water and alcohol, and showing in aqueous solution soluble easily
[a] D
=+18.8
(c
1.5,
no mutarotation observable at
this dilution).
1-Ribose gives the ordinary reactions of the pentose sugars. Reduction with sodium amalgam gives the inactive pentite alcohol adonite, which has been found in nature in the juice of the plant Adonis
Tests.
vernalis.]
1-ribonic lactone
Oxidation of 1-ribose with bromine gives 1-ribonic acid ([a] D = 18.0) and with nitric acid inactive ribotrioxycharacteristic
glutaric acid.
The most
165 C.
hydrazine derivative
is
1-ribose-bromoto
phenylhydrazone which consists of
colorless crystals melting at 164
1-Ribose-phenylosazone is identical with that of 1-arabinose, as lows from their configuration.

d, 1-Ribose.
fol-
natural adonite
Racemic ribose is formed by the oxidation by means of bromine.

2
of
CH OH HCOH 2 HCOH + HCOH CH OH

2
CHO CH OH HCOH HCOH = HCOH + HCOH + 2 H O HCOH HCOH CH OH CHO

2 2
2
Adonite
d-Ribose
1-Ribose
The sugar is also formed by molecular rearrangement from d, arabinose by means of dilute alkalies.
d, 1-Ribose
1-
has not as yet been resolved into
its optically
active
constituents.
Pentose Sugars of
Unknown
Character and Constitution.
In
addition to the pentose sugars just described different investigators have reported from time to time a number of other pentose sugars, the
*
Chem.
Centralbl. (1908), II, 1584;
(1909), II, 14.
Podwyssotzki, Archiv Pharm. (1889), 141.
560
isolation
SUGAR ANALYSIS
and
identification of
which
still
remain a matter of doubt.
Among
such pentoses*
may
be mentioned:
Cerasinose, found by Martin in the hydrolytic products of cherry
gum.
Prunose, found
by Garros in the hydrolytic products of plum gum. found by O'Sullivan in the hydrolytic products of Traganthose, Michaud
in the hydrolytic products of
gum tragacanth. Cyclamose, found by

bulbs.
It
cyclamen
can be shown upon purely theoretical grounds that no other aldopentose sugars can exist than the eight forms already described, viz.,
d-
and
1-arabinose, d-
and
1-xylose, d-
and
1-lyxose
and d- and
1-ribose.
This
may
2
be seen from the following classification:

d-xylose
d-arabinose
d-lyxose
d-ribose
CH OH
CH OH
2
CH OH
2
CH OH
2
HOCH HOCH HCOH CHO

l-arabinose
HCOH HOCH HCOH CHO

l-xylose
frOCH
HCOH HCOH CHO

l-lyxose
HOCH HOCH HOCH CHO

l-ribose
CH OH
2
CH OH
2
HCOH HCOH HOCH CHO
HOCH HCOH HOCH CHO
CH OH HCOH HOCH HOCH CHO

2
CH OH HCOH HCOH
2
HCOH CHO
The possibilities of stereo-isomerism in the aldopentoses are exhausted by the above eight forms, and the existence of new undiscovered
sugars in this class is, therefore, precluded. The sugars of unknown character just mentioned either belong to some one of the known pentoses or else fall in another class.
KETOPENTOSES
None of the ketose sugars belonging to the pentose group has as yet been isolated although several of them have been prepared in an impure form. The number of normal ketopentose sugars theoretically
*
See Lippmann's " Chemie der Zuckerarten," p. 154, for a
fuller
account of
these doubtful sugars.
THE MONOSACCHARIDES
possible
is
561
only four; in the ketose class the group adjoining the is a ketone CO replaced by aldehyde group, while the CHO is itself replaced by a CH2 OH group. It can be seen, therefore, from the preceding classification that d-araboketose is the ketone derivative of d-arabinose and d-ribose; 1-araboketose is the ketone derivative
HCOH
CHO
of 1-arabinose
and 1-ribose; d-xyloketose is the ketone derivative of and d-xylose 1-lyxose; 1-xyloketose is the ketone derivative of 1-xylose and d-lyxose.
d-Arabbketose.
CH OH
2
HOCH HOCH
U
CH OH
2
urine of rabbits fed

of ferrous sulphate.
This sugar has been detected together with d-arabinose in the It has also been prepared synupon d-arabite.* thetically by oxidation of d-arabite f with hydrogen peroxide in presence
d-Araboketose gives not only the furfural reaction of the but also the reactions characteristic of the ketoses. It forms an osazone with methylphenylhydrazine, melting at 173 C., thus differing from the aldose d-arabinose. With phenylhydrazine it forms an osazone which is identical with those of d-arabinose and dribose (as is necessary from their configuration).
Tests.
pentoses
1-Araboketose.
CH OH
2
HC(
H COH
C=
This sugar has been detected by Neuberg t in the oxidation products
of 1-arabite.
Tests.
and the resorcin and other reactions characteristic

phenylosazone
*
1-Araboketose gives the furfural reaction of the pentoses of the ketoses. Its
is
identical with those of 1-arabinose
and
1-ribose (as is
necessary from their configuration).

Neuberg and Wohlgemuth,
Ber., 34, 1745.
J Z. physiol. t Neuberg, Ber., 35, 962. Chem., 31, 564.
562
d, 1-Xyloketose.
SUGAR ANALYSIS
-
CH OH
2
THE MONOSACCHARIDES
A
riboketose
563
fall
must from
its
configuration necessarily
in the
araboketose
class.
METHYLPENTOSES
RHAMNOSE.
Isodulcite.
Rhamnodulcite.
CH
CHOH HCOH HOCH HOCH C HO

This sugar occurs widely distributed in nature as a vegetable glucosides. The latter are condensation products of sugars with alcohols, aldehydes, phenols, acids, oils, resins, alkaloids and other substances; the glucosides are hydrolyzed by acids, and also in most cases by specific enzymes, with liberation of the sugar and other constituents of the glucoside molecule. The follow-
Occurrence.
of
component
many
ing glucosides are
named out
of a large
number -which
yield
rhamnose
upon hydrolysis:
The rhamnose,
from
Quercitrin* a dyestuff obtained from the bark of Quercus citrina. or isodulcite, sold on the market is nearly all made
this source.
C2l!l220i2
Quercitrin
+ HO =
2
CeHÔs
H
5
-f-
CisHioOy.
Quercetin.
Rhamnose hydrate
Frangulin^ a dyestuff from the bark of Rhamnus frangula.
C
In
iH 20
2H O = C H
2
12
-H O
2
Frangulin
Rhamnose hydrate
is
Ci 6 HiqO 5
Emodin.
many
cases a second sugar
associated with rhamnose as a con-
stituent of the glucoside, as
Sophorin,^ a glucoside from Chinese yellow berries.
C 27 H 30
Sophorin
16
+ 3H
= CH
6
12
-H
12
+ C H O
15 10
7.
Rhamnose hydrate
d-Glucose
Sophoretin.
Hesperidin,
27
a glucoside found in
2
many
plants of the Aurantiacece.

d-Glucose
Hesperetin.
+ 3H
=
Rhamnose hydrate
Hesperidin
In the case of glucosides which are hydrolyzed into several sugars, the latter probably exist in the original compound as a higher saccharide;
*
Rigaud, Ann., 90, 283. Schwabe, Chem. Ztg., 12, 229.
J Forster, Ber., 16, 215.
Will, Ber., 20, 1186.
564
SUGAR ANALYSIS
has in fact been isolated.
in several cases this higher saccharide
Thus
xanthorhamnin, C34H 42 O 2o, a glucoside obtained from Rhamnus sagrada and other plants, when hydrolyzed by the enzyme rhamninase, gives a crystalline trisaccharide sugar rhamninose;* the latter upon heating with dilute acids is hydrolyzed into 2 molecules of rhamnose and 1 molecule of galactose (see page 731).
C H 32
18
14
4H
2C H
6
12
-H2
+ CH
6
12
6.
Rhamninose
Rhamnose hydrate
d-Galactose
The best material for the preparation of rhamnose Preparation. the commercial quercitrin or xanthorhamnin. The material is hydrolyzed with dilute sulphuric acid in the same manner as described for
is
and 1-arabinose. The acid solution is then neutralized by means barium carbonate and the filtrate evaporated to a sirup under diminished pressure. The sirup thus obtained will deposit crystals of rhamnose hydrate; the yield may be increased by precipitating gums and other impurities from the sirup by means of alcohol. The sugar is
1-xylose
of
purified
by
recrystallization.
Rhamnose exists in two forms; as rhamnose hydrate Hi 2 O 5 H 2 O and as rhamnose anhydride CeHÔs. Rhamnose Hydrate. The common crystalline form of rhamnose
Properties.
consists of large beautiful crystals having a sweetish taste but leaving an after-sensation of bitterness. The melting point of this form of rhamnose is given by different observers from 70 C. to 110 C, a circumstance due to the disturbances produced by the evolution of the water of crystallization. The constant specific rotation of rhamnose
hydrate is [a]^ =+8.5; the value decreasing somewhat with increase in temperature (see p. 179). The [a]|j 2 minutes after solution is this value diminishes and after about 10 minutes [a] D = 0; the 5;
becomes dextrorotatory attaining the constant value about one hour. +8.5 Rhamnose Anhydride. Rhamnose hydrate begins to give up its water of crystallization at 70 C. the water is completely removed by drying in a thin at 100 to 105 C., when rhamnose anhydride the sugar layer is obtained as an amorphous vitreous mass. By pulverizing the latter and dissolving it in hot water-free acetone, Fischer f obtained the anhydride in the form of white needles melting at 122 to 126 C. The constant specific rotation of rhamnose anhydride is which [a]*J =+9.4, to the of the for its water of corrected corresponds cryshydrate [<x]|j tallization. The value for [a] D of rhamnose anhydride one minute aftei
rotation then
in
;
solution
*
is
+31.5 (Fischer
J).
f Fischer, Ber., 28, 1162. J Ber., 29, 325.
Tanret, Compt. rend., 129, 752.
THE MONOSACCHARIDES A peculiarity observed in the case of rhamnose is that solution is levorotatory; [a] D constant in ethyl alcohol =
Tanret
*
its
565
alcoholic
9.0.
has explained the peculiar mutarotation of rhamnose hydrate and anhydride by the existence of several isomeric forms.
Rhamnose is not fermented by yeast; certain bacteria, however, bring about destructive changes with production of acetic, lactic and other acids.
Tests. Rhamnose reduces Fehling's solution and gives all the other general reactions characteristic of the reducing sugars. It also gives the group reactions of the methylpentoses, giving methylfurfural
upon
distillation
with hydrochloric acid.
Reduction of rhamnose
C 6 Hi 4 05, which [a] D = -f 10.7. Oxidation of rhamnose with bromine produces rhamnonic acid, whose lactone, C 6 Hi 05, shows [a] D of about 39.
with sodium
amalgam produces
the methylpentite rhamnite,
for
FUCOSE.
CH CHOH HOCH HOCH HC(OH CHO

3
,'
Occurrence. Fucose has not been found free in nature, but its mother substance, a methylpentosan (fucosan), is widely distributed in the vegetable kingdom. Fucosan has been found by Tollens and Widtsoe f in seaweed (varieties of Fucus, whence the name fucose), Irish moss, many vegetable gums and other plant materials. It has also been found by Tollens and Oshima % in "Nori," a Japanese food product prepared from the purple laver (Porphyra laciniata), and seems to be almost universally Fucosan upon hydistributed as a constituent of the marine algae.
drolysis with dilute acids
is
converted into fucose
(CH 3 C 5 H
-
4 )n
+ nH O =
2
CH C H
3 5
5.
Fucosan
Fucose
Preparation.
Tollens.
Fucose
is
One
is
kilo of dried
best prepared according to the method of seaweed (washed as free as possible from
cut into fine pieces and then treated in the cold for 24 hours with 2 per cent sulphuric acid in order to dissolve mineral salts and other impurities. The seaweed is then pressed, washed several times
sand, etc.)
with cold water and then hydrolyzed with 6

*
liters of
5 per cent sulphuric

Ber., 33, 132.
Compt.
rend., 122, 86.
t Ber., 33, 132.
% Ber., 34, 1422.
566
SUGAR ANALYSIS
The hot acid extract is then acid for 8 hours in a boiling-water bath. carbonate calcium and filtered. The with neutralized pressed out,
then evaporated to a sirup in presence of a little calcium The sirup is purified by precipitating gums, etc., with The alcohol and the alcoholic solution evaporated to a second sirup.
filtrate is
carbonate.
process of purification by means of strong alcohol is again repeated; the final sirup obtained from these purifications is then treated in the After 24 hours the fucose-phenyl'hydracold with phenylhydrazine. has which out, is filtered off, and recrystallized crystallized zone,
from dilute alcohol. The final product should be nearly white and should melt at about 170 C. The fucose-hydrazone is then decomposed with benzaldehyde (p. 348)
5 parts hydrazone, 5 parts benzaldehyde, 5 parts alcohol and 4 parts water for one-half to 1 hour upon the water bath in a flask connected with a reflux condenser. After cooling, the solution is filtered from benzaldehyde-hydrazone, shaken out with ether, clarified with
by heating
animal charcoal and evaporated to a sirup. The latter is set aside in a cool place to crystallize; crystallization can be hastened greatly by priming the sirup with a minute crystal of fucose from a stock prepaAfter crystallization is complete the sugar is filtered off (or ration.
dried
upon an unglazed
plate)
and
recrystallized.
1
The
yield of fucose
by
of
this
procS
is
3 to 8 grams from
kg. of seaweed.
Fucose consists of microscopic needle-shaped crystals Properties. an agreeable sweet taste, and easily soluble in water. The sugar shows in aqueous solution a constant rotation of about [a] D =75.5. 124. The rotation immediately after solution exceeds Fucose gives all the reactions characteristic of the methylTests. pentoses, such as production of methylfurfural upon distillation with hydrochloric acid and the color and spectral reactions described on p.
hydrazine derivatives the phenylhydrazone melting at p-bromophenylhydrazone melting at 181 to 183 C., and the diphenylhydrazone melting at 198 C. are among the most characteristic. Oxidation of fucose with bromine gives fuconic acid,
384.
its
Of
171
C., the
the lactone of which,
HioO 5 gives a rotation

,
of
[a]
RHODEOSE. -
=+
78.3.
Cu
HCOH HCOH HOCH
mo
THE MONOSACCHARIDES
Occurrence.
free in nature; it occurs,
567
Rhodeose, the antipode of fucose, has not been found however, the same ft its isomer, rhamnose, as a constituent of certain vegetable glucosides. The best-known glucoside in which rhodeose has been found is convolvulin,* the purgative
Convolvulin upon hydrolysis principle of jalap (Convolvulus purga). and a acid mixture convolvulinic of sugars consisting of gluyields
and isorhodeose; it is supposed that these sugars are united in the glucoside to form a complex saccharide. Rhodeose is best prepared from the commercial v Preparation.
cose, rhodeose
convolvulin; 50 gms. of convolvulin are dissolved in 375 c.c. of bariumhydrate solution (saturated at room temperature). The excess of ba-
rium is precipitated by carbon dioxide and sulphuric acid, and the filtrate, which should contain exactly 0.5 per cent free sulphuric acid, heated The filtered solution is then neutralized with to 100 C. for 40 hours. barium carbonate and clarified with 5 c.c. of saturated lead-subacetate The filtrate is freed from lead by means of hydrogen sulphide, solution. and the filtered solution evaporated under reduced pressure to a sirup, which contains the sugars in the proportion of 2 parts of rhodeose to 1 part of glucose. The glucose is removed from the sirup by fermentation
with yeast; the rhodeose
ylhydrazine as
is
is
then precipitated by means of methylphen-
The latter after recrystallizing several times with benzaldehyde and the decomposed by warming filtered solution after shaking out with ether evaporated to a sirup,
an insoluble hydrazone.
which
is then set aside in a cool place for crystallization. Priming the with minute a a of rhodeose from sirup crystal previous preparation will hasten crystallization.
Rhodeose consists of small needle-shaped crystals a sweet taste and easily soluble in water. The sugar shows in having solution a constant rotation of [a] D = +75.5 (+86.5 after soluaqueous
Properties.
tion).
Tests.
Rhodeose gives
ylpentoses.
in
As the
the reactions characteristic of the methoptical antipode of fucose it resembles the latter
all
The diphenylhydrazone of its behavior with many reagents. rhodeose melts at 199 C. (that of fucose at 198 C.); the p-bromophenylhydrazone of rhodeose melts at 184 C. (that of fucose at 183 C.); the methylphenylhydrazone of rhodeose melts at 174 C. (that of fucose at 177 C.), etc. Upon oxidation with bromine rhodeose gives rhodeonic acid, the salts of which have the same composition as those of
fuconic acid.
*
The
of fuconic acid at 106 to 107
lactone of rhodeonic acid melts at 105.5 C. (that C.); in their rotatory power, however, the
I,
Taverne, Chem. Centralbl. (1895),
56; Votocek, Ber., 37, 3859; 43, 469.
568
SUGAR ANALYSIS
[a] D
two lactones show their antipodal character, = -76.3 (lactone of fuconic acid = +78.3).
rhodeonic acid lactone
This racemic comRacemic Sugar from Rhodeose and Fucose. was prepared by Votocek * by evaporating a solution containThe sugar was obtained as ing rhodeose and fucose in equal amounts. minute crystals melting at 161 C. and optically inactive; it is much
bination
less soluble in
water than either of
its
components.
Isorhamnose.
CH
CHOH
HCOH HOCH HCOH CHO
This methylpentose has not been found in nature; it has been prepared synthetically by reduction of the lactone of isorhamnonic acid (made
by heating rhamnonic acid and pyridine

Properties.
to 150 C.). Isorhamnose has not been isolated as yet in the crystalline form; as prepared by Fischer and Herborn f the sugar was obtained as a sweet easily soluble sirup which showed a value for [a] D of about -30. Tests. Isorhamnose gives methylfurfural upon distillation with hydrochloric acid and gives the other reactions of the methylpentoses. Oxidation with bromine gives isorhamnonic acid, the lactone of which shows after solution [a] D = 62, which however sinks in 24 hours to about 5, owing to decomposition of the lactone into free acid. The phenylosazone of isorhamnose is identical with that of rhamnose; this of course follows necessarily from the structural relationship of the two
sugars.
Quinovose.
Occurrence.
CH C H
3 5
5.
This methylpentose has been found as a constituent of the glucoside quinovin, which occurs in the bark of different varieties of the cinchona tree.
Preparation.
Quinovin upon hydrolysis with hydrochloric acid in
alcoholic solution yields quinovose, which in presence of the alcohol and acid is converted into ethyl quinovoside, C2 5 This com5 6
C HnO
Ber., 37, 3859.
t Ber., 29, 1961.
THE MONOSACCHARIDES
pound,
first
569
called quinovite,
hydrolysis, until Fischer
was long regarded as a direct product of and Liebermann * ascertained its composition
and showed
it
to be the result of a secondary reaction.
The
ethyl
quinovoside upon heating 1| hours with 3 parts of 5 per cent sulphuric The solution is diluted acid is hydrolyzed into alcohol and quinovose.
1 vol. of water, the alcohol evaporated, the acid neutralized with barium carbonate and then, after decolorizing with bone black, the liquid The filtrate is extracted with ether to remove any unhydrofiltered. and then evaporated, when the quinovose is obtained quinovoside lyzed
with
as a yellowish sirup.
Quinovose has been obtained only as a yellowish Properties. sirup of strong dextrorotation, easily soluble in water and absolute alcohol.
amounts
Quinovose reduces Fehling's solution, and yields large upon distillation with 12 per cent hydrowhen heated with phenylhydrazine give quinovose-phenylosazone which consists of fine yellow needles melting at 193 to 194 C. When heated with hydrochloric acid in alcoholic soluTests.
of methylfurfural chloric acid. Its solutions
tion quinovose gives the ethylglucoside previously referred to. Ethylquinovoside is an amorphous hygroscopic substance melting at 60 C.
and when pure should be perfectly soluble = is dextrorotatory, [oi] D +78.1.

Isorhodeose.
in ether.
The compound
This methylpentose of unknown f together with rhodeose among The sugar has been obtained the hydrolytic products of convolvulin. dextrorotation of low as a +20 about). ([a] D only yellowish sirup
3
5
CH C H
5.
configuration was found by Votocek
Isorhodeose-phenylosazone consists of fine yellow crystals melting at 190 C.

i
SUGARS OF
UNKNOWN
CONSTITUTION, ISOMERIC OR RELATED TO THE
METHYLPENTOSES
Antiarose. C 6 Hi2 5 This sugar was obtained by Kiliani t in the hydrolysis of the glucoside antiarin which is found in the sap of the upas tree (Antiaris toxicaria), the milky juice of which is used by Malayans as an arrow poison. Antiarose has only been obtained as a
.
sirup; it gives the reactions of an aldose and, oxidized with bromine, = 30. 5 is levorotatory, [a] D yields antiaronic acid whose lactone, C 6 Hi
,
Ber., 26, 2415. t Ber., 43, 476.

j
Archiv. Pharm., 234, 438.
570
SUGAR ANALYSIS
C?!^^. This sugar, discovered by Kiliani,* is Digitalose. It is formed by the hydrolysis of to be a dimethylpentose. supposed the glucoside digitalin, which is found in different species of digitalis.
C 5H
3
56
Oi4
+ 2H
Digitalin
= CaHp, Digitaligenin
C7H 14
Digitalose
H O + 2 H O. + CGlucose
6
12
of
Digitalose has been obtained only as a sirup and gives the reactions an aldose sugar. Oxidation with bromine gives digitalonic acid C 7 Hi 4 6 whose lactone is levorotatory [a] D = 79.4.
,
,
HEXOSES
ALDOHEXOSES
D-GLUCOSE.
sugar, etc.
Dextrose.
Grape sugar.
^^j CHjOH
Starch sugar.
Diabetic
lL HOC] /V^-tl
HOCH HCOH
HOCH
Occurrence. d-Glucose is the most widely distributed sugar in it is in the free condition in the blood and tissues of found nature;
most animals, and in the juices of nearly all plants. The sugar occurs most abundantly, however, in the combined form in such substances as the vegetable glucosides, the higher sugars and the polysaccharides.
The Vegetable Glucosides.f The reactivity of the glucose molecule in nature is best exemplified by the vegetable glucosides, condensation products of glucose with alcohols, acids, aldehydes, phenols and other compounds. Reference was made to several glucosides
which yielded rhamnose upon hydrolysis under the description of the latter sugar. The glucosides, which contain glucose as their sugar constituent, are,
impossible to describe
* t
however, the most widely distributed in nature. It is all of these, but a few typical examples are given.
Ber., 26, 2116; 31, 2454.
For a
full
see the section
account of the various glucosides, their preparation, properties, etc., by Euler and Lundberg in the Biochemisches Handlexicon (1911),
Vol. II, pp. 578-722; also Armstrong's "Simple Carbohydrates and Glucosides" " Chemical Changes and Products resulting from Fermenta(1910) and Plimmer's " tions (1903).
THE MONOSACCHARIDES
The
glucosides are usually colorless crystalline taste, and for the most part levorotatory.
Salicin.-
571
bitter
compounds with a
a remedy for rheumatism.
glucoside found in the bark of the willow and used as It is hydrolyzed by the enzyme emulsin to
glucose and salicyl (o-oxybenzyl) alcohol.
13
18
+ H
= C
fl
12
+ CH
6
(OH)CH OH.
2
Salicin
Glucose
Salicyl alcohol.
glucoside found in the bark of several species of poplar (Populus). It is hydrolyzed as follows:
Populin.
C20 H
22
+ H
A
= CH
6
12
+ C H COC H
6
(OH)CH2 OH.
trees.
Populin
Glucose
fir
Benzoyl-aalicyl alcohol.
Coniferin.
It is
glucoside found in the follows: as hydrolyzed
and other coniferous
C H22
16
+ H
.=
CH
6
12
Coniferin
Glucose
Coniferyl alcohol.
Arbutin. This glucoside together with methylarbutin is found in the leaves of the evergreen bearberry (Arbutus uva ursi). The two glucosides are hydrolyzed as follows:
12
16
+H
2
= C
12
+CH
6
(OH) 2
Arbutin
Glucose
Hydroquinone.
Ci 3 Hi 8 O7+
Methylarbutin
HO =
CfHaOi
Glucose
CeH 4 Qjj
Methylhydroquinone.
A glucoside found in the bark of the apple, pear and Phloridzin. other trees belonging to the Rosaceae. It possesses the peculiar property of causing glucosuria when taken internally; the amount of glucose
in the urine
may reach from

It
6 per cent to as high as 13.5 per cent after

is
ingestion of phloridzin.
hydrolyzed by acids as follows:

2
C iH
2
2 4Oio -f-
HO =
CBHÔG
Glucose
-f-
CuHuOk*
Phloretin.
Phloridzin
Gauliherin.
lyzed by
A glucoside found in the wintergreen. acids as follows:

-
It is
hydro-
Ci 4
Indican.
18
+H
= C 6 Hi 2
Glucose
+ C H OHCOOCH
6
3.
Gaultherin
Methyl
salicylate.
lyzed by
acids or
glucoside found in the Indigo plant. by the enzyme indimulsin as follows:
It is
hydro-
C H N0 + H
14 17
6
= C 6H 12
Glucose
+CH
6
/ X
NH
^CH COH
Indican
Indoxyl.
572
SUGAR ANALYSIS
of the
The indoxyl
above reaction
is
colorless,
but undergoes rapid
oxidation to indigotin the blue coloring matter of indigo.
2C H
6
NH X ^>CH+O
/
= C
/
4
NH
COH
^CO
>C:C
NH
^CO
>C H + 2H O.
6 4 2
Indoxyl
Indigo-blue.
glucoside found in the root of the madder It is hydrolyzed by a specific (Rubia tinctorum) and other plants. enzyme or by acids into glucose and the coloring substance alizarin.
Ruberythric
Add.
14
+2H
CH
6
12
+CH
6
/ '
4
co \
C 6H (OH) 2
2
Ruberythric acid
Glucose
Alizarin.
A glucoside found in bitter almonds and in the kerplums and other fruits of the same family. Amygdalin was the first glucoside to attract investigation; it was discovered in 1830 by Robiquet* and 7 years later Liebig and Wohlerf discovered the manner in which it was hydrolyzed by emulsin, an enzyme found with amygdalin in the almond. Amygdalin is the most interesting of the glucosides not only historically but also from the peculiarity of
Amygdalin.
nels of peaches,
giving off hydrocyanic acid
upon
hydrolysis.
CaoHsAiN
Amygdalin
+ H
2
CH
6
12
+ C H CHO + HCN.
6
5
Glucose
Benzaldehyde
Hydrocyanic
acid.
It
has been supposed that the two glucose molecules in amygdalin
are united to form a disaccharide.
The
5
HCN
in
the nitrile of 1-mandelic acid
C H CH(OH) CN. The

6
amygdalin occurs as monoglucose
compound
of 1-mandelonitrile
was obtained by Fischer by hydrolyzing

.
amygdalin with an enzyme found in yeast; it has the following formula, C 6 H 5 CH(CN) - O - C 6 Hn0 5 This glucoside is also found in nature in the bark of the wild cherry and other trees. There are a large
family of glucosides belonging to the mandelonitrile class. Besides amygdalin and its derivative 1-mandelonitrile glucoside, the following
are mentioned:
A glucoside found in the leaves of the common Sambunigrin. (Sambucus nigra). It is the monoglucose compound of d-mandelonitrile, the optical antipode of the derivative obtained by Fischer t from
elder
amygdalin.
It is
17
hydrolyzed by acids and emulsin as follows:

6
C 14 H
laurel.
N+H
A
= CH
6
12
C H CHO
6 5
HCN.
acid.
Sambunigrin
Glucose
Benzaldehyde
Hydrocyanic
Prulaurasin.
It is
* t
glucoside found in the leaves of the cherry a racemic mixture of d-mandelonitrile glucoside (sam[2],
Robiquet and Boutron, Ann. chim. phys.

Ann., 22, 1-24 (1837).
44, 352-382 (1830).
J Ber., 28, 1508.
THE MONOSACCHARIDES
bunigrin) and 1-mandelonitrile glucoside. way as sambunigrin.
It is
573
hydrolyzed in the same
and
A glucoside found by Dunstan and Henry* in the leaves It is a para-hydroxymandelonitrile Sorghum vulgar e. glucoside and is hydrolyzed by emulsin and acids as follows = C 6 Hi2 6 + C 6 H 4 (OH)CHO + HCN. Ci 4 H 17 7 N + H 2
Dhurrin.
stalks of
:
Dhurrin
Glucose
p-Oxybenzaldehyde
Hydrocyanic
acid.
growth
by eating sorghum due to the hydrocyanic acid derived from this glucoside. A glucoside found by Dunstan and Henry Phaseolunatin. Lfrna beans (Phaseolus lunatus), flax and cassava (also termed
is
The poisoning
of cattle
at certain stages of its

in
lini-
marin)
It yields the following hydrolytic
products
3
10
17
N+H
= C
12
Phaseolunatin
Glucose
COCH + + CHAcetone
3
HCN.
acid.
Hydrocyanic
Sinigrin. glucoside found in the seed of black mustard (SinaIt is one of the most interesting of the glupis or Brassica nigra). cosides, as it contains sulphur and yields a mustard oil as one of its
hydrolytic products.
called
it
The
glucoside
was
first
isolated
by Bussy who
potassium myronate. It is hydrolyzed by acids or by the enzyme myrosin (which accompanies it in mustard seed) as follows:
Ci Hi 6 O 9 NS 2
Sinigrin
K+H
= C Hi
6
Glucose
NCS + KHS0 + CH :CHCH Potassium mustard

2
4.
Allyl
oil
bisulphate
glucosides constitute a separate class and are peculiar to the plants of the Cruciferae. Sinalbin. This is the glucoside of the white mustard (Sinapis or
The mustard-oil
Brassica alba).
It is
hydrolyzed by the enzyme myrosin which ac-
companies
it
in the following
2 S2
manner:
12 6 7
7
C3oH 42
O N
15
+H
= C
Sinalbin
H O + C H ONCS + Ci H O N HS0 Sinalbin mustard Glucose Sinapin

6
24
4.
acid-sulphate.
oil
Glucose
is
also
found associated with tannic and
gallic acids as
constituent of another very widely distributed group of plant subThese have been regarded by some chemists as true glustances.
cosides
and by others as mere complexes. Glucotannin, a so-called glucoside of this class, hydrolyzed by the enzyme tannase as follows:
is
supposed to be
C 27 H 22
The
state that
17
+4H
= C
12
+ 3 CH
(OH) ? COOH.
Glucotannin
Glucose
Gallic acid.
literature
no
definite
*
upon the tannin glucosides is in such a contradictory compounds can be cited by way of illustration.
Phil, trans.
t Proc.
Roy.
Roy. Soc., 1902, Soc., 72, 285.
199, 399.
574
SUGAR ANALYSIS
The glucosides are best isolated from Preparation of Glucosides. plant materials by extraction with alcohol. The extraction should be begun as soon as possible after the material is gathered in order to prevent hydrolysis by accompanying enzymes. In many cases the glucoside will crystallize directly from the alcohol extract; in other cases, where the yield is small, the extract must be concentrated before cryslarge amounts of other organic substances are present, clarification with lead salts may be necessary, in which case any excess of lead is afterwards removed by means of hydrogen
tallization will begin.
When
sulphide.
Besides forming Glucose as a Constituent of Higher Sugars. condensation products with plant alcohols, aldehydes, phenols, acids, cometc., to form glucosides, glucose may unite in other ways; it may molemore or with one or bine with one or more molecules of itself, The cules of other sugars, to form the higher crystalline saccharides.
latter are readily
specific
hydrolyzed into the component sugars by acids and

in the
enzymes
same manner as the
glucosides.
The
following
examples are given of the higher sugars found in nature which yield
glucose
upon
hydrolysis.
Sugar
Hydrolytic Products
Maltose Trehalose
Disaccharides,
Gentiobiose Sucrose
Turanose
Lactose Melibiose
Trisaccharides,
Melezitose Gentianose
Raffinose
+ glucose. + glucose. + glucose. + fructose. + glucose. (?) + galactose. + galactose. Glucose + glucose + glucose. (?) Glucose + glucose + fructose. fructose + galactose. Glucose
Glucose Glucose Glucose Glucose Glucose Glucose Glucose
-j-
Tetrasaccharides, C24H42O21-
Stachyose
Glucose
+ fructose + galactose + galactose.
In addition to Glucose as a Constituent of Polysaccharides. own its of condensation molecule, glucose forming higher sugars by may unite with itself to form the very complex polysaccharides, such as
The number of molecules of dextrin, dextran, starch and cellulose. these higher derivatives is which enter the structure of into glucose
* indicate Researches by Brown and Morris that dextrin, the simplest member of the class, contains 40 molecules of glucose in the condensed form and that the molecule of starch is at least 5 times as large as that of dextrin, in other words contains 200
difficult to
determine.
Chem.
Centralbl. (1890),
I,
845.
See also
Brown and
Millar, J.
Chem.
Soc.,
76, 315.
THE MONOSACCHARIDES
575
molecules of glucose. Cellulose, the most highly condensed polysaccharide, has without doubt a molecule much greater even than starch.
The chemical properties of the higher polysaccharides can only be referred to very briefly. Cellulose. (C 6 Hi O 5 )n. This is the most abundant constituent
forms probably dry vegetable matter of the world. Cellulose occurs in the pure condition only in the fiber of the cotton ball and in a few similar substances; as a constituent of the cellular tissue of plants cellulose is usually combined with lignin, pentosans and other hemicelluloses to form a complex of varying composition. By treatment of plant membranes with hot solutions of dilute alkalies and
in the vegetable
kingdom; makes up one-half of the
cellulose in all its various
total
acids the lignin, pentosans and other so-called encrusting sub" are split off from the complex leaving the cellulose behind as stances
"
The latter upon bleaching with chlorine is obtained in a perfectly white fibrous form. The average approximate percentage of cellulose in the water-free substance of different plant
an insoluble residue.
materials
is
as follows:
Material (water free)
Approximate per cent
of Cellulose
Wood
Bark Straw
Leaves Seeds (including husks) Roots, tubers, etc
60 40 40 20
15 10
Cellulose, as prepared from its different sources, consists of a white fibrous material insoluble in ordinary solvents but easily soluble in an ammoniacal copper solution. It is reprecipitated from the latter by
acids as a gelatinous mass. Cellulose swells up in concentrated sulphuric acid and after short contact is converted into a starch-like sub-
In the same manner stance (amyloid) which is colored blue by iodine. cellulose gives a blue coloration with zinc chloride and iodine solution
Cellulose after long contact with concentrated (Schulze's reagent). sulphuric acid is dissolved; upon diluting the mixture with water and boiling the cellulose is hydrolyzed to d-glucose. Starch. (C 6 HioO 5 ) 2oo. (?) Starch is next to cellulose the most
widely distributed glucose
condensation product
in
the
vegetable
up as a reserve material in many roots, tubers, grains and seeds, in which parts it may constitute over 90 per cent of * that 10 the total dry substance. It has been estimated by Nageli per cent of the Phanerogams or flowering plants produce starchy seeds.
kingdom.
It
is
stored
"Die Starkekdrner"
(1858), p. 378.
576
SUGAR ANALYSIS
starch-yielding capacity of certain plants, as the cereals, potato, yam, cassava, etc., is so pronounced as to give them great value in the production of starch for food and industrial purposes. The oc-
The
currence of starch in the leaves and chlorophyll tissue of plants has al-
ready been referred to. Starch is deposited in plant
whose
192).
size
cells in the form of minute grains, and shape are peculiar to each botanical species (see Fig.
By
reducing the starchy parts of the plant to a fine pulp with
Wheat
starch.
Rice starch.
Corn
Fig. 192.
starch.
Potato starch.
Forms
of starch grains.
After Moller.
(Magnified 400.)
water and straining the milky liquid into a tall cylinder the grains of starch will be deposited as a sediment. The latter is then washed several times by decantation with 0.25 per cent sodium hydroxide solution, which removes protein and other impurities, and then washed with water to remove all traces of alkali. The starch thus prepared
and dried at a gentle heat. In a similar way, by a process which is largely mechanical, starch is prepared commercially from the potato, cassava, Indian corn, wheat, rice and other plants. Pure starch, which has not been subjected to heat or strong chemiis
filtered off
form of white microscopic granules varyThe granules ing in size from about 0.002 mm. to 0.2 mm. diameter. are insoluble in cold water, but when heated with hot water burst and partially dissolve forming a thick mucilaginous paste.
cal treatment, occurs in the
The conversion
under maltose.
of starch
by means
of diastase
and acids
is
is
described
Starch upon contact with cold hydrochloric acid
converted into
THE MONOSACCHARIDES
soluble starch.
577
is
In Lintner's
method the
starch
treated for 7 to 8
days with 7.5 per cent hydrochloric acid, after which it is washed free from acid and dried. The product thus obtained dissolves readily in hot water to form a clear limpid solution. The [a] D of soluble starch 195 to 200. ranges from about Dextrin. (C 6 Hio0 5 )4o. (?) Dextrin is formed in nature and in the
by the conversion of starch. It occurs in malted grain and in all starchy seeds during germination. The dextrin molecule is regarded by many chemists as varying in character and the following classificaarts
tion
is
sometimes made.
Amylodextrin,
first dextrin of conversion; blue iodine reaction. Erythrodextrin, second dextrin of conversion red iodine reaction. (3) Achroodextrin, third dextrin of conversion; no iodine reaction. (4) Maltodextrin, final dextrin of conversion; no iodine reaction.
(1)
(2)
have
subdivisions of dextrins under each of the above groups been proposed. The theories concerning the formation of dextrin from starch are discussed under maltose.
also
Numerous
stable dextrin was prepared by Brown and Millar f by precipitating a diastatic starch conversion with alcohol, dissolving the precipitate in water, fermenting away occluded sugars with yeast, and again After several such purifications and fracprecipitating with alcohol. tional precipitations with alcohol, a dextrin was obtained of only slight
reducing power and giving for
[a]
195 to
+ 195.7.
The formula
39(C 6 Hio05)
assigned to this dextrin (see p. 687). Dextrin is prepared commercially by heating starch with about The temperature of heating varies ac0.2 per cent nitric acid in ovens. cording to the color and character of product desired, and ranges from
G
C Hi20 was
6
110
is
to 170
C.
from starch by
dextrin thus prepared resembles that obtained It is easily soluble in water, and diastatic conversion.
The
The specific precipitated from aqueous solution by strong alcohol. to the method of rotation of commercial dextrins will vary according
manufacture
Such dextrins, purified by repeated precipi(see p. 510). tation with alcohol, will give a value for [a] D of about +195, the same as that of the stable dextrin prepared by diastase. Dextrin upon heating with dilute hydrochloric or sulphuric acid is
hydrolyzed into d-glucose. The reaction is not perfectly quantitative, however, owing to a slight destruction of the glucose. The rate of hydrolysis was found by W. A. Noyes | and his co-workers to be only about onehalf that for maltose.
*
J.
prakt. Chem., 34
t J.
[2],
381.
t J-
Chem.
Soc., 76, 315.
Am. Chem.
Soc., 26, 266.
578
SUGAR ANALYSIS
In addition to the Other Glucose-yielding Polysaccharides. substances previously mentioned glucose occurs in the condensed form in many other plant products, such, for example, as lichenin* (C 6 Hio0 5 ) n a constituent of many mosses and lichens; dextran (C 6 Hio0 5 ) n a muci,
,
laginous substance secreted

its
properties; paradextran^
fungi other materials belonging to the so-called hemicelluloses. In the animal kingdom glucose is found free in the blood (about 0.1 per cent) and also in very slight amounts in normal urine (usually
by many bacteria and resembling dextrin in (CeHuAOn, a cellular constituent of many and mushrooms; plant dextrins and gums, vegetable-glycogen,^. and
under 0.01 per cent or less than 0.5 gm. per day). In diabetes mellitus and other diseases where glucosuria occurs, the per cent of glucose in the urine may exceed 10 per cent with a daily excretion of 500 or even 1000 gms. Temporary glucosuria may be produced by ingestion of phloridzin (p. 571), various alkaloids, potassium chlorate and other
compounds.
Glucose occurs most abundantly in the animal kingdom in the form
of its condensation product glycogen.
is a constituent of all growing animal regarded as a reserve product, playing the same role in the animal economy as starch in the vegetable. The surplus
Glycogen.
(CeHnAOn
cells.
It is usually
one-half of this being deposited in the liver and other organs.
carbohydrates of the food are stored up in the body as glycogen, about and one-half in the muscles
The percentage
of glycogen in fresh
meat and muscles
varies from a
trace to nearly 3 per cent; it is found in largest amount in the liver where it may constitute over 10 per cent of the weight of this organ.
The amount
of glycogen in the liver is subject to considerable fluctuation being greatest after a meal rich in carbohydrates and lowest after It is only after several weeks' fasting, long abstinence from food.
however, that glycogen disappears completely from the liver. In its removal from the various organs of the body glycogen is hydrolyzed by enzymes to glucose, which is then transported by the blood to the different parts of the organism.
Preparation of Glycogen. Glycogen is prepared by cooking finely divided livers with 60 per cent potassium hydroxide solution for two
* t
Bauer,
J.
prakt Chem.,
[2]
34, 46.
Winterstein, Ber., 27, 3113.
j Errera,
Compt.
rend., 101, 253.
Discovered by Claude Bernard in 1855 (Compt. rend., 41, 461; 44, 1325; 48, 884, etc.). For a full account of this carbohydrate consult Pfluger's book " Das Glykogen," 2nd. Edit. (1906), Bonn.
THE MONOSACCHARIDES
hours.
579
The filtrate is then diluted to 15 per cent potassium hydroxide and, after settling, the clear solution is mixed wit]?. 1 volume of 96 per The precipitated glycogen is washed with a mixture of 1 cent alcohol. volume of 15 per cent potassium hydroxide, and 2 volumes of 96 per cent
alcohol. The crude product thus prepared is purified by dissolving in strong potassium hydroxide solution and reprecipitating with alcohol. Glycogen is obtained as an amorphous snowProperties of Glycogen.
white substance. The product is usually more or less hydrated so that it is difficult to prepare a substance of uniform composition. Glycogen dissolves in water to a faintly opalescent colloidal solution; it is also dissolved by hot alcohol the best solvent is aqueous potassium hydroxide
;
Gly cogen is strongly dextrorotatory, [a] D = about +200. It is decomposed by butyric acid bacteria and other organisms, but is not fermented by yeast. Plant enzymes of different origin convert glycogen, some into maltose and some into d-glucose; the enzymes of the body, however, seem to hydrolyze glycogen only into d-glucose. Glycogen is hydrolyzed by acids into d-glucose in the same manner as starch, dextrin, and other polysaccharides of the glucose class.
solution.
Glycogen does not reduce Fehling's solution. Treated with iodine it gives a color varying from brown to wine-red, which disappears upon heating to 60 C. but returns again upon cooling. Of other animal products, which may be regarded as glucose derivatives, may be mentioned tunicin* or animal cellulose (C 6 Hio0 5 ) n found in the outer membranes of Tunicates and other animals, and the socalled animal gum found by Landwehr f. in various tissues and organs
solution
,
of the body.
A number of animal substances, which are The Gluco-proteids. not of a pure carbohydrate nature, yield glucose as one of their hydrolytic Among these substances may be mentioned different nuproducts. The cleo-proteids, various albumins and certain mucins or mucoids.
chemistry of these products, however, is still unsettled, and it is uncertain whether the sugar derived from them consists of glucose alone
or of a mixture of sugars.
Honey.
Another animal product rich
in glucose
is
honey, although
the sugar in this instance is primarily of vegetable origin being derived by bees and other insects from the nectar of flowers and other plant
juices.
Honey
contains usually about 35 per cent of glucose; strained
honey
*
will frequently granulate
and even solidify owing to

227.
crystallization
Berthelot,
Compt.
Chem.,
rend.,
47,
Winterstein,
Z.
physiol,
Chem.,
18,
46-56.
f Z. physiol.
8.
122;
9,
367; 18, 193; 19, 339; 20, 249.
580
of its glucose.
SUGAR ANALYSIS
The granulation
of
crystallized glucose as thus observed known to mankind.
and
honey was known to the ancients, was probably the first sugar
d-Glucose has been synthetised in a Synthesis of Glucose. It has been prepared from d-mannose, by oxidizing of ways. this sugar to d-mannonic acid and converting the latter by molecular rearrangement (p. 775) into d-gluconic acid, whose lactone upon reduc-
number
The sugar has also been built up synthetically of formaldehyde to d, 1-fructose, which condensation by by and this upon oxidation d, 1-man1-mannite reduction d, gives upon nonic acid. The latter is then resolved into its d- and 1-components and d-glucose is prepared from the d-mannonic acid as first described.
tion gives d-glucose.
Fischer *
d-Glucose can be prepared directly Preparation of Glucose. from granulated honey by stirring the mass with a little 50 per cent The sugar thus obtained is alcohol and filtering upon a suction plate. dextrin and other from fructose, honey impurities by recrystalpurified
lization.
its
In preparing glucose upon a large scale hydrolysis of some one of natural condensation derivatives is employed. For this purpose starch, the raw material from which commercial glucose is made, is
usually chosen.
One hundred grams of pure Preparation by Hydrolysis of Starch. potato or corn starch are heated to boiling with 1000 c.c. of 2 per cent hydrochloric acid in a flask connected with a reflux condenser for 2
hours.
and
filtered.
The hot solution is neutralized with lead carbonate, cooled The filtrate is evaporated to a thin sirup, shaken with an
equal volume of hot 96 per cent alcohol and filtered from precipitated The alcoholic filtrate upon evaporation gives a sirup which impurities.
rapidly crystallizes; the sugar thus obtained
crystallization.
is
purified
by a second
Glucose can also be prePreparation by Hydrolysis of Cellulose. pared by hydrolysis of cellulose; 100 gms. of clean cotton or filter paper are slowly stirred into 500 c.c. of 80 per cent sulphuric acid. The mixture is allowed to stand 24 hours; it is then diluted to 5000 c.c. and heated in a boiling water bath for 5 hours. The hot solution is then neutralized with an excess of calcium carbonate, filtered and the filtrate concentrated to a sirup. The latter is then purified from gummy decomposition products by heating with alcohol through bone black. The alcoholic filtrate
and clarified by filtering upon evaporation gives a
sirup which soon crystallizes.

*
Ber., 23, 799.
THE MONOSACCHARIDES
581
Glucose is also very easily prePreparation by Inversion of Sucrose. pared by inversion of cane sugar. For this purpose 1000 gms. of refined sugar are heated for 1 hour with 300 c.c. of water and 5 c.c. of concentrated hydrochloric acid in a flask immersed in a boiling water bath. The yellowish colored sirup is then poured into an evaporating dish and set aside in a cool place. Granulation will begin after a few weeks' standing
crystals.
filtered,
when the glucose will separate as a thick or even solid mass of The latter are taken up with a little 50 per cent alcohol,
washed with strong alcohol and
recrystallized in the usual
way.
The
its
crystallization of glucose can always be hastened by priming sirups with a small crystal of glucose from a previous prepara-
tion.
Glucose crystallizes both as the hydrate Properties of Glucose. Hi206.H 2 O and the anhydride. The hydrate is obtained by crystallizing from water at ordinary temperature and the anhydride by Glucose hydrate crystallizing from a hot saturated alcoholic solution. loses its water of crystallization between 50 and 60 C. To prepare an-
hydrous glucose from the hydrate, the latter should be spread in a thin layer in a flat bottomed dish and heated at 60 C. until most of the water has been expelled; the temperature is then gradually raised to
100 C. when the sugar will be obtained perfectly granular and dry. Heating the hydrate directly to 100 C. will cause melting and when this occurs it is difficult to obtain a satisfactory preparation of anhydrous
glucose.
146
Glucose anhydride consists of fine crystalline needles melting at to 147 C. The sugar has a sweet taste, is very soluble in water (about 1 1 at 20) and hot alcohol, but insoluble in ether. The con:
stant rotation of the anhydride is [a]D 106 or more). For glucose hydrate
= 4-52.5
[a] D
(directly after solution
=+48.2
(constant).
Tan-
has considered glucose to exist in 3 modifications, a high-rotating form (H D =+106), a constant-rotating form ([a^ =+52.5) and a low-rotating form ([a] D =+22.5). Tanret's constant-rotating glucose is now considered to be simply an equilibrated mixture of a-, or highReference has been rotating, glucose and /?-, or low-rotating, glucose.
ret
made
to these modifications under the subject of mutarotation. Glucose undergoes a large number of Fermentations of Glucose. fermentations a few of the more typical of which will be mentioned. Glucose is fermented to alcohol by many Alcoholic Fermentation.
species of Saccharomyces,
*
Mucor, Torula, Mycoderma and other organrend., 120, 1060.
Compt.
582
isms.
SUGAR ANALYSIS
The
given by Gay-Lussac C 6 Hi 2 O 6 = 2
Glucose
was
first
general formula for the fermentation of glucose to alcohol * as follows:
CH CH OH + Alcohol
3 2
C0
2.
Carbon
dioxide.
or,
51.11 parts alcohol +48.89 parts carbon dioxide. Pasteur, f however, showed that this formula was not absolutely correct, as he obtained under the best of conditions only 48.3 parts of alcohol and
100 parts glucose
46.4 parts of carbon dioxide from 100 parts of glucose. Pasteur obtained in addition to alcohol and carbon dioxide 2.5 to 3.6 parts of glycerol, 0.4 to 0.7 part of succinic acid and 1.3 parts of fat, cellulose and
other substances, all of which he believed to be formed directly from the sugar. The latest researches, however, show that these minor products of fermentation are the result of metabolic processes within the yeast cells. The main phase of the fermentation is produced by the
enzyme zymase which is secreted by the yeast. Buchner,| in fact, has separated zymase from crushed yeast and by its aid has fermented glucose into alcohol and carbon dioxide without the direct agency of
the living
cells.
Fig. 193.
Saccharomyces
cerevisise.
After Hansen.
In the alcoholic fermentation of glucose a concentration of 5 to 20 per cent sugar gives the best results. The temperature should be maintained between 30 and 35 C., and a small amount of peptones, phosphates, tartrates and other salts be added as nutrient
for the yeast.
which
is
A growth of Saccharomyces cerevisice, or beer yeast, one of the best known alcohol-producing organisms, is shown
in Fig. 193.
In addition to ethyl alcohol a number of other alcohols are pro*

t
Ann. chim. phys. Ann. chim. phys.

"
[1],
95, 311.
68, 330, 355, 362.
[3]
t Ber., 30, 117, 1110,
2668; 34, 1523,
etc.
See also the book by Buchner and
Hahn
Die Zymasegarung," Munich, 1903.
THE MONOSACCHARIDES
583
duced in the alcoholic fermentation of glucose; the most important of these are amyl, isobutyl and propyl alcohols with traces of hexyl alcohol and higher homologs. These higher alcohols (the fusel oil of distilleries)
* according to the researches of Ehrlich are not formed from the
sugar, however, but are secondary products derived from the acids of the yeast.
amino
R CHNH COOH + H
2
= R
Amino
CH OH +
2
CO
acid
Higher alcohol
Carbon dioxide
NH
3.
Ammonia.
Lactic Fermentation.
Glucose
is
fermented to
lactic acid f
by a
The formula for the fermentalarge tion of glucose into lactic acid in its simplest terms is expressed as follows:
of bacilli
number
and
bacteria.
CH
6
12
CH CHOH COOH.
3
d-Glucose
Lactic acid.
is never obtained, more carbon dioxide, hydrogen, formic, acetic, butyric and propionic acids, mannite and other products of secondary or foreign fermentative origin being always formed. In fermenting glucose to lactic acid a 10 per cent solution of the
The
theoretical yield of lactic acid, however,
or
less
prepared (adding minute amounts of ammonium salts, nibran extract or other substances to serve as nutrients) and then trates, sterilized. The cold solution is inoculated with a pure culture of Basugar
is
other lactic acid producing organism) and incubated C. for 3 to 6 days. A little powdered calcium carbonate is added from time to time to take up the excess of free acid which should be kept below 0.5 per cent, but never be entirely neutralized (otherwise
cillus lactis acidi (or
at 45
to 55
the butyric fermentation may set in). If the lactic acid culture is pure, 98 per cent of the glucose may be converted into lactic acid by
this
method.
The lactic acid obtained by fermentation is usually optically inactive and consists of a racemic mixture of the d- and 1-isomers. Ceror
tain organisms have been found, however, which seem to form the d-, In these cases d, 1-lactic acid may per1-, acid alone or in excess.
haps be formed first, the d-, or 1-component being afterwards partly or wholly destroyed by secondary fermentation. The conversion of d-glucose into lactic acid, according to Buchner,t is due to the action of a special enzyme secreted by the organisms. Glucose is fermented into butyric acid Butyric Fermentation.
by a number
*
of species of bacteria;
among
the best
known
of these is
Ber., 40, 1027-47.
t Ber., 36, 634.

[3], 2,
t Pasteur,
Ann. chim. phys.
257 (1842).
Pasteur,
Compt.
rend., 62, 344.
584
SUGAR ANALYSIS
the Clostridium butyricum. The butyric fermentation, which proceeds only in the absence of air, follows approximately the following equation:
Hi 2
= CH CH CH COOH
3 2 2
Glucose
Butyric acid
CO + 2 H + 2Carbon
2
Hydrogen.
dioxide
As by-products of the butyric fermentation there are generally formed butyl, ethyl and propyl alcohols, formic, acetic, propionic, valeric, caprylic, capric and lactic acids, and other substances either of secondary or foreign fermentative origin. The butyric acid producing bacteria grow best at a temperature between 35 and 40 C. and require a medium perfectly free from acid.
To
secure the latter condition the butyric fermentation
is
carried out
in presence of an excess of calcium carbonate free acid as fast as it is formed.
which combines with the
Viscous Fermentation.*
of different
Glucose
is
organisms to a mucilaginous
fermented by a large number gummy substance known as
The gum which is formed during fermentation 'is probably a secondary product, being a constituent of the gelatinous capsule which encloses the organism. Among the best-known organisms,
dextran.
which produce the viscous fermentation, is the Leuconostoc mesenterioides f (Fig. 196) to which class belong also the Bacterium pediculatum J (Fig. 197) and other gum-forming organisms found in sugar factories. The bacteria which form gum thrive best in the absence of air, and at a temperature varying from 30 to 35 C.
Dextran^, the gum of the viscous fermentation, and the cause of " ropiness in wine and of the so-called frog-spawn" in sugar factories, is precipitated from solution by means of alcohol. It is purified by
dissolving in dilute sodium hydroxide, filtering and reprecipitating with alcohol acidified with acetic acid. The product, after drying and pulverizing, is obtained as a white powder, having the general formula
The gum
(CeHioOs)^ and yielding glucose upon hydrolysis with sulphuric acid. swells up in water but does not form a perfectly translittle alkali.
parent solution except upon addition of a

strongly dextrorotatory, [a] D = about Oxidizing Fermentations of Glucose.
Dextran
is
fermentations of glucose,
* t j
+200. There are a large number of which differ from the types previously de30, 32. Centralbl., (1894), II, 703. dextran see references in Lippmann's
Pasteur, Bull. soc. chim. (1861), 30.
Van Tieghem, J. fabric, sucre, 20, Alfred Koch and Hosaeus, Chem.
For the copious literature upon der Zuckerarten," 427-430.
"Chemie
THE MONOSACCHARIDES
scribed, in that different acids.
585
oxygen is absorbed from the air with the formation of Such fermentations belong to the strictly aerobic class.
d-Glucose is fermented to d-gluoblongus,* Bacterium xylinum\ and nuIn its simplest phase the reaction proceeds
The
following are examples: Gluconic Acid Fermentation.
conic acid
by the Micrococcus
merous other organisms.

as follows:
CH OH
2
CH OH
2
(CHOH) 4
(CHOH) 4
CHO
d-Glucose
COOH
d-Gluconic acid.
With some organisms the fermentation proceeds almost quantitatively

according to this reaction. In other cases the gluconic acid itself undergoes oxidation so that the theoretical yield is much reduced.
Citric
Acid Fermentation.
Citromyces group ferment glucose into for the reaction is given as follows:
Various organisms belonging to the citric acid. The general formula
CH OH
2
(CHOH)
+ 3O
CHO
Glucose
CH - COOH C(OH) - COOH CH - COOH

2
2
+2HO
2
Citric acid.
attended as the formula shows by a of about 200 c.c. of oxygen being taken up high consumption oxygen, from the air for every gram of glucose fermented. In fermentation experiments the yield of citric acid from glucose
citric acid
The
fermentation
is
has not been found to exceed 50 per cent, owing to the fact that the
citric
acid itself undergoes oxidation in the later stages of the fer-
mentation.
Glucose is fermented into oxalic acid The complete conversion a number of moulds bacteria. and by large of glucose in this direction, however, is never reached, as the oxalic acid at a certain point of concentration begins to exercise a toxic effect
Oxalic Acid Fermentation.
upon the growth of the organisms. The oxalic acid, which is formed by the action of microorganisms, is probably a respiration product given off by the living cell rather than a true fermentation derivative (such as alcohol) formed from the sugar by the action of a special enzyme. The same is no doubt also true of many other acids. Glucose is subject to a large number Other Acid Fermentations.
*
Boutroux, Compt. rend., 91, 236.
Brown,
J.
Chem.
Soc., 49, 432; 60, 463.
586
of other fermentations in
SUGAR ANALYSIS
which
acetic, propionic, formic, succinic
and
other acids are formed.
In some of these cases the fermentation seems
to be of a true enzymic character. Certain bacteria, for example, are able to convert glucose directly into acetic acid.
CH
6
12
CH COOH
3
Glucose
Acetic acid.
in certain fermentations
Buchner and Meisenheimer * have been able to show that this change is produced by a specific enzyme which they
found possible to
isolate. Many investigators believe that all other acid fermentations of glucose are also the result of special oxidizing enzymes or oxidases; this view, however, requires considerable more experimental proof before it can be accepted.
Ester Fermentation. In many fermentations of glucose, as by the Bacillus suavolens, the production of alcohol and acids proceeds simultaneously, the result being the formation of different fruity esters, such
as ethyl acetate, ethyl butyrate and ethyl valerate. The chemistry of the different compounds, which are formed in the
fermentation of glucose and other sugars, is so extensive that the student is referred for further particulars to the special works upon
The products obtained by upon d-Glucose. with caustic alkalies have been briefly referred to. heating d-glucose The chief product of this decomposition is d, 1-lactic acid, the occurrence of which in molasses is due to the action of lime during clarification upon glucose and fructose. The lactic acid is easily obtained from molasses by acidifying with sulphuric acid and extracting with ether.
the subject.f Action of Alkalies
Another alkali-decomposition product of glucose, found in sugar-house products, is saccharin, C 6 Hi O 5 the lactone of saccharinic acid C 6 H 12 6 Saccharin is prepared according to
Saccharin.
is
which
Scheibler by dissolving 1 kg. of d-glucose (or d-fructose) in 7 to 8 liters of water, and adding, with continuous boiling, freshly slacked milk of lime, so that the solution is still alkaline after 3 to 4 hours. The cooled
solution
is
saturated with carbon dioxide and the filtrate treated with
oxalic acid just sufficient to
combine with
is
all
the lime.
The
filtrate
containing the saccharinic acid

Saccharinic acid, which
*
evaporated to a sirup, long standing deposits crystals of saccharin.

is
which after
an isomer of glucose,
is
the a-methyl
Ber., 36, 634.
t Particularly
Lippmann's
"
Chemie der Zuckerarten " and
Lafar's
"
Tech-
nische Mykologie."
THE MONOSACCHARIDES
derivative of d-arabonic acid, whose structural formula saccharin are as follows
:
587
and that
of
CH OH HOCH HOCH H C-COH COOH

2
3
Saccharinic acid (a-methyl-d-arabonic acid)
CH OH CH HOCH +HO
2
2
CO
Saccharin (a-methyl-d-arabonic
acid lactone).
Saccharin forms clear rhombic double-refracting crystals which melt It is easily soluble in at 160 C. and sublime without decomposition. is not fermented and alcohol ether, by yeast, and does not rewater,
duce Fehling's solution. The specific rotation is [a] D = +93.5. Saccharin is a type of a large group of isomeric substances * (isosaccharins, metasaccharins, etc.) which are produced by the action Nef by the action of 8 normal sodium of alkalies upon different sugars. a- and 0- dextro-metasaccharins. obtained hydroxide upon d-glucose
CH OH HOCH CH HCH HOCH

2
CH OH HOCH . -- CH HCH HCOH

2
I
a-Dextro-metasaccharin
[a]
j8-Dextro-metasaccharin
D = +25.28
m.p.
[lz>
104 C.
m.p.
= +8.2 = 92 C.
No single sugar has been subjected to Tests for d-Glucose. such a variety of tests as d-glucose, and the various compounds which have been obtained in its combination with different reagents number
many
hundred. Glucose in its reduction of alkaline solutions of copper,
silver,
mer-
cury and other metals, in its color reactions and many other tests behaves in the same way as other sugars of the aldose class. The following reactions are given as
among
the
more
characteristic of glucose
and
its
derivatives.
Glucose and the various subwhich stances yield glucose upon hydrolysis are oxidized by strong
Saccharic Acid Test for Glucose.
*
701, 2953; 16, 2625;
For further particulars upon the different saccharins see Kiliani 18, 631, 2514) and Nef (Ann., 376, 1).
(Ber., 15,
588
SUGAR ANALYSIS
which
test,
is recognized by means of its acid according to Tollens and Gans,* is
nitric acid to saccharic acid,
potassium or silver salt. The best carried out as follows:
Five grams of the material to be examined are treated in a porcelain dish with 30 c.c. of nitric acid of 1.15 sp. gr. and the mixture evaporated with constant stirring upon a boiling water bath until evolution of
red fumes has ceased and the resulting sirup has just begun to take on a permanent yellow color. The sirup is then taken up with a little
water, heated over a flame and powdered potassium carbonate added until a drop of the brownish colored solution gives a blue reaction with
red litmus paper. Glacial acetic acid is then added drop by drop until the mixture gives off a strong odor of acetic acid. If glucose was present in the original substance crystals of acid potassium saccharate will
usually soon separate; if crystallization does not take place after a few hours' standing, as may happen when only small amounts of glucose are present, the sirup should be concentrated further by gentle evapoAfter 24 hours' standing the crystals, which have formed, are ration.
drained upon unglazed porcelain, and then recrystallized from the smallest possible amount of hot water. A third crystallization, using bone black, will usually eliminate the last traces of oxalic acid and other impurities and give a perfectly pure salt. The yield of acid potassium saccharate by this method is about 30 to 40 per cent of the
filtered off, or
amount of glucose. The compound consists of shining rhombic crystals with characteristic trapezoidal faces, the appearance of Acid potassium sacwhich under the microscope is unmistakable.
original
charate has the formula
COOH (CHOH) COOK.

4
acid potassium saccharate as above prepared can be further For this purpose the acid identified by conversion to the silver salt.
The
potassium
to which
tion
is
salt, after
is
ammonia
drying and weighing, is dissolved in a little water, then added to the point of neutrality. The solu-
then poured into a cold silver nitrate solution containing amount of 1 \ the weight of the acid potassium salt taken. The precipitated saccharate of silver after standing a short time is filtered, washed free from silver nitrate and then dried in a dark place over concentrated sulphuric acid. The silver saccharate has the for-
AgNOa
to the
mula CeHsOsAga and upon
ignition in a porcelain crucible should show 50.91 per cent Ag. In making the saccharic acid test for glucose it should be remem-
bered that d-gluconic, d-glucuronic and d-gulonic acids and d-gulose also give saccharic acid upon oxidation with nitric acid. This limita*
Ber., 21, 2149.
THE MONOSACCHARIDES
tion,
589
action
however, is a comparatively slight one and the saccharic acid reupon the whole is one of the best tests for d-glucose in presence
d-Glucose forms a large num-
of other sugars.
Hydrazones and Osazones of Glucose.
ber of hydrazones with phenylhydrazine and its substituted derivatives. With phenylhydrazine itself two isomeric hydrazones are
formed, one modification melting at 144 to 146 C. and the other at 115 to 116 C. The exact conditions under which these two isomers
are formed
(see p. 359). Fischer *
and
their structural relationship are not fully understood
and Stahel
recommended the diphenylhydrazone
as one
of the best
compounds for identifying glucose. To carry out the test 1 part of sugar is dissolved in as little water as possible and 1.5 parts of
diphenylhydrazine in alcoholic solution are added; if the mixture is not clear a little water or alcohol is added until the turbidity disappears.
the mixture to stand several days the diphenylhydrazone N(C6H 5 ) 2 will separate as small colorless prisms, after to which, recrystallizing from hot water, should melt at 161 162 C. By heating the mixture of sugar and diphenylhydrazine in a
6
By allowing CH N
12 5
:
flask attached to a reflux
condenser the formation of the hydrazone
is
completed within 2 hours. Upon evaporating the alcohol and taking up the uncombined hydrazine with ether the hydrazone will quickly The glucose-diphenylhydrazone is insoluble in ether and separate.
addition of the latter will hasten crystallization. The diphenylhydrazone reaction offers an easy means for separation of d-glucose from d-fructose and other sugars. In case of a mixture of sugars it is advisable to conduct the reaction in the cold. It should be remembered
in
making the
test that arabinose also gives a characteristic insoluble
hydrazone with diphenylhydrazine; the two compounds can easily be separated, however, by crystallization and are readily distinguished from one another by their differences in melting point and composition. COCeH5 and methylGlucose-benzhydrazone J C 6 Hi 2 O5 N NCH3 C 6 H5 have also been emCeHÔs: N phenylhydrazone ployed for the separation and identification of glucose. Glucose may be separated from its hydrazones by decomposing the latter with benzalThe free sugar, dehyde or formaldehyde as described on page 348. which is thus liberated, may be crystallized and further identified by
:
NH
determining
*
its specific
rotation.
earliest studied
The best-known and

Ber., 23, 805. t Ann., 258, 242.
hydrazine derivative of glucose

t Wolff, Ber., 28, 160.
Neuberg, Ber., 36, 965.
590
SUGAR ANALYSIS
:
C 6 5 ) 2 The compound is is the phenylosazone C 6 Hi O 4 ( obtained as yellow needle-shaped crystals by heating 1 part glucose, 2 parts phenylhydrazine chloride and 3 parts sodium acetate in 20 The osazone is purified by recrystallizing from alcohol, parts of water.
.
NH
and should have a melting point

method). The osazone reaction of glucose
of 205
to 206
C. (capillary tube
is of but little value as a means of owing to the fact that d-mannose and d-fructose both give the same compound. d-Glucose upon treatment with sodium Reduction of Glucose. amalgam in acid solution is reduced to the alcohol d-sorbite.
identification,
CH OH
2
THE MONOSACCHARIDES
591
The a- and /3-methyl glucosides, although having the same general formula, C 7 Hi 4 O 6 show a marked difference in certain of their properties, as is seen from the following:
,
592
1-Glucose.
SUGAR ANALYSIS
CH OH
\-/
Hi OH
HCOH HOCH HCOH
1-Glucose has not been found free in nature

theoretical interest
and possesses only a
from the relationship to its d-isomer. The sugar has been prepared synthetically * from the pentose sugar 1-arabinose. The latter upon treatment with hydrocyanic acid and saponincation of the addition product (Kiliani's cyanhydrine synthesis) gives a mixture of the two hexonic acids.
CH OH
2
HCOH HCOH HOCH HCOH

l-Gluconic acid.
CqOH
CH OH HCOH HCOH HOCH HOCH COOH

2
1-Mannonic
acid.
The 1-gluconic and 1-mannonic acids are separated tion of their lactones. The lactone of 1-gluconic acid sodium amalgam to 1-glueose.
Properties.
by
is
crystalliza-
reduced by
ing at 141
d-glucose.
to 143
1-Glucose consists of small prismatic crystals melting C. In its outward properties the sugar resembles
Its specific rotation
was found by Fischer
to be
[a] D
51.4;
mutarotation was present, the reading after solution being 95.5. 1-Glucose is not fermented by yeast and in this respect shows an
important difference from d-glucose.
sium and
Nitric acid oxidizes 1-glucose to 1-saccharic acid whose acid potassilver salts resemble those of .d-saccharic acid.
Diphenylhydrazine forms with 1-glucose a characteristic hydrazone, which melts at 162 C. and is in other respects very similar to d-glucose-diphenylhydrazone. The phenylosazone of 1-glucose is formed under the same conditions as d-glucose-phenylosazone and resembles the
latter in melting point, crystalline
*
form and
solubility;
it is
readily dis-
Fischer, Ber., 23, 2611.
THE MONOSACCHARIDES
tinguished, "however, from the d-glucose-osazone in glacial acetic acid solution.
593
by
its
dextrorotation
Racemic glucose was obtained by Fischer,* as a d, 1-Glucose. sweet colorless inactive sirup, by dissolving equal parts of d- and 1-glucose in water and evaporating.
d, 1-Glucose gives with diphenylhydrazine a colorless diphenylhydrazone of melting point 132 to 133 C., which is 30 C. below the
melting point of either d- or 1-glucose-diphenylhydrazone.
Oxidation
of d, 1-glucose with nitric acid gives d, 1-saccharic acid, whose acidpotassium salt is very similar to those of its d- and 1-components.
Racemic glucose is easily resolved by means of yeast, the d-glucose being completely fermented and the 1-glucose remaining behind.
D-MANNOSE.
Seminose.
CH OH
2
HOCH HOCH HCO H HCOH

CHO.
Occurrence.
d-Mannose
is
found in the free condition accord-
ing to different investigators in the juices of various plants and in many germinating seeds. The sugar has also been found in certain molasses
from tropical cane-sugar factories; the mannose in molasses, however, not derived from the cane but is formed by the action of the lime used in clarification upon the glucose and fructose of the cane juice. d-Mannose is most widely distributed in nature as an anhydride condensation product mannan which in its simple or complex form makes up one of the most abundant of the hemicelluloses. Mannan (C 6 Hi O 6 ) n This carbohydrate, either in its simple form or as one of the so-called " paired mannans" (glucomannan, galactomannan, fructomannan, etc.), occurs in nearly all plants from the simplest unicellular Protophytes to the most highly developed Phanerogams. It is found in the cellular matter of yeast, in different moulds, in various alga, in plant gums, in the wood, bark and roots of many trees, and in bulbs, nuts, grains, seeds, fruits, berries, leaves and
is
.
other plant tissues.

*
594
Yeast
SUGAR ANALYSIS
Mannan.
convenient material for preparing one of the mannans is pressed yeast (manufactured without starch). Salkowski's * method of separating and purifying yeast mannan, or yeast gum, is as follows: 500 gms. of yeast are boiled gently for one-half hour with 5 liters of 3 per cent potassium hydroxide. The solution is then al-
lowed to stand, and the clear liquid, which is poured off, heated with 750 c.c. of Fehling's solution upon a water bath. The yeast gum is thrown out as a bluish white insoluble copper compound, which is purified from its mother liquor by boiling and squeezing out with water. The copper compound is then decomposed by rubbing up with a slight excess of hydrochloric acid and 4 to 5 volumes of 90 per cent alcohol added. The acid alcoholic solution is poured off from the precipitated gum; the latter is then dissolved in water and reprecipitated by After washing with a little absolute alcohol and alcohol as before. in 25 parts of water, a few drops of the is redissolved ether, gum acid the solution poured into 7 volumes are added and hydrochloric of absolute alcohol. The precipitated gum is washed with alcohol and ether and then allowed to remain under ether until the plastic mass hardens to a brittle solid. The ether is then poured off, the gum ground up in a mortar and dried over sulphuric acid. Yeast mannan consists of a white non-hygroscopic powder, easily soluble in warm water to a clear limpid solution and showing a specific rotation of 90.1. Hydrolysis of yeast gum with hydrochloric Z) =Hor sulphuric acids gives large amounts of d-mannose. According to
Oshima d-glucose and fucose are also formed, so that yeast gum in all probability belongs to the complex mannans. A very pure mannan has been prepared from the Salep Mannan.
mucilage of salep, a drug consisting of the dried decorticated tubers of different orchidaceous plants. Salep yields from 40 to 50 per cent of mucilage; the latter when separated from insoluble matter conalmost wholly of a water soluble mannan, which can be precipitated from solution by means of alcohol. Salep mannan f has the general formula (C 6 HioO 5 ) n and is hydrolyzed by acids first to lower mannosaccharides and finally to d-mannose. Mannans have been isolated from many other plant substances,
sists
the general method of preparation consisting in the extraction of the material with hot 2 to 3 per cent alkali and precipitation of the gum with Fehling's solution as described under the method for yeast gum.
The combining power of
Fehling's solution with

*
mannan is very marked,
Ber., 27, 497, 925.
t Hilger, Ber., 36, 3198.
THE MONOSACCHARIDES
the reagent causing a precipitation of
liquid.
1
595
in 1000 parts of
part
mannan
Preparation of Mannose.
Mannose may be prepared
either
by
hydrolysis of yeast gum, salep mucilage or any other of the isolated mannans, or by direct hydrolysis of some plant rriaterial rich in mannan.
The
latter
method
is
being several
common
the most direct and the easiest to carry out, there vegetable substances, such as ivory nuts, carob
beans, coffee berries, date seeds, etc., which yield large amounts of mannose upon hydrolysis. Ivory nuts, or vegetable ivory (the fruit of Phytelephas macrocarpa), which is used so extensively for making buttons,
is
of Fischer
one of the best substances for preparing mannose. and Hirschberger * is as follows
:
The method
One part of ivoryPreparation of d-Mannose from Ivory Nuts. nut shavings (from button factories) is heated with 2 parts of 6 per cent hydrochloric acid in a boiling water bath for 6 hours in a flask connected with a reflux condenser. The hot solution is then pressed out from the insoluble residue and the latter treated with a little water and repressed. The combined extract is then neutralized with sodium hydroxide, decolorized with bone black, filtered and treated in the cold with an excess of phenylhydrazine (0.3 gm. for every 1 gm. of
ivory-nut shavings) dissolved in acetic acid. The very insoluble mannose-hydrazone soon separates as a thick crystalline precipitate, which
is filtered off
after 24 hours, washed with cold water and dried. The hydrazone may be purified by recrystallizing from a large volume of hot water or from 50 per cent alcohol containing a little pyridine, but for the purpose of preparing mannose the troublesome recrystallization may be dispensed with. Mannose may be separated from its hydrazone by any of the methods
previously described (p. 348). The best procedure according to Tollens f is the following: 50 gms. of mannose hydrazone, 40 gms. of benzaldehyde, 50 gms. of alcohol and 50 gms. of water are heated upon the
water bath for about one-half hour in a flask connected with a reflux condenser. The solution is then cooled and filtered from the insoluble benzaldehyde-hydrazone; the filtrate is shaken out with ether, deThe colorized with bone black, filtered and evaporated to a sirup.
latter rarely crystallizes until it has
been primed with a crystal of mannose from a previous preparation. Crystallized mannose has been prepared by van Ekenstein J by dis*
Ber., 22, 3218.
t j
" Biochem. Arbeitsmethoden Abderhalden's Rec. trav. Pays-Bas, 14, 329; 15, 222.
"
(1909), II, 74.
596
SUGAR ANALYSIS
solving mannose sirup in methyl alcohol, adding a half volume of ether and after 24 hours decanting from the sirupy precipitate. The
methyl-alcohol-ether solution on long standing will deposit crystals of
mannose, which may be used for priming impure sirups. After the mannose-containing sirup has crystallized, the sugar is freed from its mother liquor by spreading upon porous plates and then
purified
by
recrystallizing.
* According to Neuberg and Mayer mannose is best separated from The sugar crystallizes at its hydrazone by means of formaldehyde. once in a highly pure condition without a trace of decomposition prod-
ucts.
d-Mannose has been Preparation of d-Mannose from Carob Beans. obtained by Herissey f from the seeds of the carob bean (St. John's bread) by allowing the mannan of the seeds to react with the accompanying enzyme, seminase: 500 gms. of the finely ground seeds are treated with a solution of 60 gms. sodium fluoride in 4000 c.c. of water and allowed to stand at 33 to 35 C. The fluoride prevents
fermentation by microorganisms, while the seminase of the seeds converts the
mannan
to d-mannose,
which
is
precipitated from solution
method described. d-Mannose has been made synthetiSynthesis of d-Mannose. d-Mannite is oxidized by dilute nitric acid to cally in a number of ways. d-mannose which can then be separated as the hydrazone. d-Mannose can also be formed from d-glucose and d-fructose, through molecular The sugar has rearrangement by action of dilute alkalies (p. 303). also been built up by Fischer J from formaldehyde; the latter by condensation gives d, 1-fructose, which upon reduction gives d, 1-mannite, and this upon oxidation with bromine yields d, 1-mannonic acid. The latter is resolved by crystallization of its strychnine salts into the dand 1-components. The lactone of the d-mannonic acid upon reduction
as the hydrazone according to the
gives d-mannose. Properties of
dride
d-Mannose. d-Mannose crystallizes as the anhyrhombic crystals melting at 132 C. The sugar has a pleasant sweet taste and is easily soluble in water and 80 per cent alcohol, very slightly soluble in hot absolute alcohol and insoluble in
Hi 2
in
ether.
d-Mannose has a constant

initial
specific rotation of [a] D
=+
14.25; the
minutes after solution being 13.6. d-Mannose is fermented by yeast to alcohol and carbon dioxide in the same manner, but not with the same The rapidity, as d-glucose.
is
rotation
to the left, [a] D 3
Z. physiol.
Chem., 37, 547.
Compt.
rend., 133, 49, 302.
J Ber., 23, 370.
THE MONOSACCHARIDES
sugar is also easily fermented by different lactic acid organisms. nose reacts with alkalies similarly to d-glucose.
597
d-Man-
Tests for d-Mannose.

of its phenylhydrazone,
d-Mannose
2
:
is
best recognized
,
by means
which repeated referO5 5 6 6 ence has been made. The compound crystallizes in colorless rhombic prisms, which melt upon slow heating at 186 to 188 C., but with rapid heating at 195 to 200 C. The hydrazone is almost insoluble in cold water, but is dissolved in 80 to 100 parts of hot water; it is but
to
little
is
C Hi
N NHC H
hot 60 per cent alcohol.
soluble in concentrated alcohol, ether or acetone; the best solvent Dissolved in hydrochloric acid it exhibits
levorotation.
On
zone,
is
long heating with phenylhydrazine, d-mannose, or its hydraconverted into an osazone which is identical with that of
(p. 354).
d-glucose
its
d-Mannose upon reduction with sodium amalgam is converted into alcohol d-mannite, which melts at 166 C. and in borax solution is
d-Mannite
is
dextrorotatory.
very widely distributed in nature and has been found
in ash
manna,
lilac leaves,
mushrooms, sea
algae
and
in the olive, cactus,
The best natural source is pineapple, onion and many other plants. ash manna, from which d-mannite is obtained by extraction with alcohol.
Oxidation of d-mannose with bromine in aqueous solution gives
d-mannonic
acid,
CH OH(CHOH) COOH,
2
whose lactone,
with
C 6 Hi
nitric
6,
is
dextrorotatory
([a] D 53.8). Upon d-mannose and d-mannonic acid give d-mannosaccharic acid, COOH (CHOH) 4 COOH, whose double lactone, C 6 H 6 6 melts at 180 to 190 C. and has a rotation of [a] D = + 201.8.
,
= -f-
oxidation
acid
1-Mannose. -
CH OH HCOH
2
I
rlOUxl
HOCH HOCH CHO

bined form;
1-Mannose has not been found in nature either it has been prepared synthetically
in the free or
*
com-
in several ways.
The
is
best starting point is 1-arabinose, which, as shown under 1-glucose, converted by means of Kiliani's cyanhydrine reaction into both The lactone of the latter upon re1-gluconic and 1-mannonic acids.
*
598
duction with sodium
SUGAR ANALYSIS
amalgam gives 1-mannose. The sugar can also be Fischer's synthesis from formaldehyde as described under prepared by the synthesis of d-mannose. 1-Mannose has been obtained only in
unfermentable levorotatory sirup. 1-Mannose forms with phenylhydrazine an insoluble hydrazone, which resembles d-mannose-phenylhydrazone in
form of a
colorless
Tests for l-Mannose.
melting point and other properties.
The 1-mannose-hydrazone, how-
ever, exhibits dextrorotation, when dissolved in hydrochloric acid, and this property serves to distinguish it from the d-mannose-hydrazone.
The phenylosazone of 1-mannose is identical with 1-glucosazone. 1-Mannose upon reduction gives its alcohol 1-mannite, which melts
at 166 C.
and in borax solution is levorotatory. Oxidation with bromine converts 1-mannose to 1-mannonic acid, whose lactone is levorotatory = ~ 54.8). Oxidation with nitric acid converts 1-mannose and ([]/>
1-mannonic acid to 1-mannosaccharic acid whose double lactone melts at 180 C. and has a rotation of about [a] D = - 200.
d,
1-Mannose.
colorless inactive
acid.
Racemic mannose was obtained by Fischer * as a sirup by reduction of the lactone of d, 1-mannonic
By decomposing d, 1-mannose-phenylhydrazone with formaldehyde Neuberg and Mayer f obtained d, 1-mannose in the form of crystals which melted at 132 to 133 C. d, 1-Mannose forms an insoluble
phenylhydrazone melting at 195 C.;
d, 1-glucosazone.
its
osazone
is
identical with
gives d, 1-mannite; oxidation with bromine yields d, 1-mannonic acid and with nitric acid d, 1-man-
The sugar upon reduction
nosaccharic acid.
the d-mannose.
1-Mannose can be resolved by means of yeast which ferments only Indirectly the sugar can be resolved by conversion to The strychnine salt of the latter is then treated d, 1-mannonic acid. with boiling alcohol which dissolves only the salt of the d-acid. The lactones of the separated mannonic acids yield upon reduction the respective sugars, d- and 1-mannose.
d,
D-GALACTOSE.
CH OH
2
HOCH HCOH HCOH HOCH

*
Ber., 23, 381.
f Z. physiol.
Chem.,
37, 545.
THE MONOSACCHARIDES
599
Free d-galactose has been reported as occurring in Occurrence. in the tissues of certain seeds during germination; milk and the whey of
the galactose thus found, however, by enzyme action from some of
is
its
purely transitory, being derived higher condensation products,
which, as glucosides, polysaccharides and hemicelluloses, are found widely distributed in nature.
d-Galactose occurs in the vegetable world as a constituent of many glucosides, in which compounds it is often united with other sugars.
The
glucoside xanthorhamnin, which gives d-galactose and rhamnose upon hydrolysis, has already been mentioned. In the same way the
is
glucoside digitonin, a constituent of commercial digitalis, by heating with dilute acids into galactose and glucose.
hydrolyzed
C27 H
40 13
+2H
= CH
6
12
Digitonin
d-Galactose
H +G +C H d-Glucose
6 12 6
24 3 . 15 Digitogenin.
Convallarin and convallamarin from Convallaria majalis (lily of the valley), myrticolorin from the leaves of Eucalyptus macrorhyncha, sapo-
from certain species of Saponaria, and many other complex glucosides, whose constitution remains to be established, yield upon hydrolysis d-galactose, which is usually mixed with other sugars. d-Galactose is also found united with other sugars as a constituent of different higher saccharides, such as lactose, melibiose, raffinose, rhamninose and stachyose. d-Galactose is found most widely distributed in the Galactans.
toxin
vegetable kingdom as a constituent of many gums, hemicelluloses, In these cases the galacmucilages, pectins and other plant materials.
tose usually exists as an anhydride condensation product or galactan. The galactans make up a numerous group of substances, the exact con-
which has not been thoroughly established. In the galactans d-galactose shows the same tendency to form combinations with other sugars as was noted in the case of its glucosides and polysacstitution of
arabogalactans, xylogalactans, mannogalactans, glucogalactans and other combinations each of which shows wellmarked differences in behavior towards alkalies and other reagents.
charides;
there are
China moss, Ceylon moss, Irish moss, Iceland moss and many other plants belonging to the algae, mosses and lichens yield mucilages, which are dissolved by hot water and precipitated therefrom by alcohol.
The substances thus prepared consist mostly of galactan and are hydrolyzed by hydrochloric or sulphuric acid to d-galactose. Oxidation with nitric acid produces large quantities of mucic acid. Arabogalactans or galactoarabans are found in the seeds of lupines,
600
beans, peas
cent.
SUGAR ANALYSIS
and other legumes
*
in
The
cellular tissues of the
amounts varying from 5 to 20 per crushed seeds are extracted succes-
sively with water, alcohol, ether and 0.2 per cent potassium hydroxide; the residue from this treatment consists largely of galactoaraban. The crude product is purified by dissolving in hot 2 per cent potassium
hydroxide and precipitating the clear solution with strong alcohol, which throws out the galactoaraban as a yellowish colored potassium compound. The latter is washed with alcohol, decomposed with dilute
and the pure gum precipitated by addition of strong alcohol. Galactoarabans have also been found in plant exudations and gums, in vegetable mucilages, in the slimy envelopes of different bacteria, in
acid
unripe sugar-beets and in

are easily hydrolyzed tose and 1-arabinose.
many other products. The galactoarabans by hydrochloric and sulphuric acids into d-galacIn
many
cases this hydrolysis can be effected
by diastase and other enzymes; the latter in the process of germination no doubt convert the galactoaraban of seeds into sugars which
are then assimilated
yield
by the growing embryo. The galactoarabans mucic acid upon oxidation with nitric acid, and furfural upon
distillation
with strong hydrochloric acid. Galactoxylan f has been found as a gummy constituent of the cellular tissues of wheat, barley and other grains, and also appears to
occur in various complex gums.

acids to galactose
Gatactoxylan is hydrolyzed by dilute and xylose. Mucic acid is obtained upon oxidation with nitric acid, and furfural upon distillation with hydrochloric acid. Galactomannan I occurs as a constituent of many hemicelluloses;
it has been found in the coffee berry, in cocoanuts, in the carob bean, in the seeds of Strychnos Ignatii (St. Ignatius' beans) and in other plant substances. Hydrolysis of galactomannan gives galactose and mannose. Galactans of a more complex character than the above are also
found in nature. Among such galactans may be mentioned flax-seed mucilage, which upon hydrolysis gives d-galactose, d-glucose, 1-arabinose and 1-xylose. The Pectins.
Closely related to the plant mucilages and
gums
are the pectins, an important group of substances widely distributed in nature. The pectins are found in apples, pears, grapes and most
*
Schulze, Ber., 22, 1192.
Lintner and Dull, Z. angew. Chem. (1891), 538. " Lippmann's Chemie der Zuckerarten," p. 694. For a fuller account of the pectins see article by Victor Grafe, " Biochemisches Handlexicon," pp. 80-94; also the early papers by Fremy to whom much of our knowledge is due (J. Pharm. [2], 26, 368 (1840); Ann. chim. phys. [3], 24, 5).
t
t
THE MONOSACCHARIDES
of
601
other fruits, in carrots, beets and other root organs, and in the tissues many other plants, as the flax and hemp.
which are soluble, are derived from an insoluble intermother substance, called pectose, which is regarded by many chemists as an oxygen, or acid, derivative of cellulose. The conversion of pectose into pectin takes place in the ripening of fruits, and in the retting of flax and hemp; the process is attributed to the action of a
pectins,
cellular
The
special
enzyme
pectosinase.
A good
juice
is
material for preparing pectin is the juice of ripe pears. The treated first with oxalic acid to break up lime compounds and
then with tannic acid to precipitate albumin. The clarified juice is filtered and treated with an excess of strong alcohol, which precipitates
the pectin.
The latter is filtered off, purified by dissolving in water and reprecipitating with alcohol, and then dried over concentrated sulphuric acid. As thus prepared pectin consists of an amorphous white substance, which dissolves easily in water to a neutral solution. The pectins differ greatly in their optical properties; the pectin from orange skins, for example, is inactive, while the pectin from gooseberries gives [a] D
=+
194.
long boiling with water pectins and pectose are converted into parapectin of weak acid reaction. Boiling with dilute acids con-
Upon
verts pectose, pectin and parapectin into metapectin, which has also the properties of a weak acid.
The pectins by the action of another enzyme pectase are converted into pectic acids, the calcium salts of which give fruit juices the propPectic acids are also produced from pectose and erty of jellifying.
pectin
by boiling several hours with dilute alkali. As precipitated from solutions of its salts pectic acid is obtained as a white amorphous jellylike mass, insoluble in water, alcohol and ether, 186 to + 300. but easily soluble in alkalies; [a] D Pectose, pectin, parapectin, metapectin and pectic acid are converted by hot alkaline solutions into parapedic acid. The final prod-
=+
uct obtained
by the action
of alkalies
upon the pectin substances

is
previously named is metapectic acid-, which acid described under 1-arabinose.
identical with arabinic
The
different pectin substances,
which have been named, are
all
hydrolyzed by boiling with dilute mineral acids into d-galactose and 1-arabinose, the yield of each sugar depending upon the nature of the
product.
hydrolysis of the pectins into galactose and arabinose is also supposed by some to take place in nature through the agency of a third enzyme pectinase.
The
602
SUGAR ANALYSIS
Oxidation of the pectins with nitric acid gives a large yield of mucic acid, and distillation with hydrochloric acid produces much
furfural.
The chemistry of the pectins is still in a very unsettled condition. The neutral water-soluble pectins are usually regarded as lactones or
esters of the various pectic acids,
but the constitution of the
latter,
as well as that of the parent substance pectose, has not been deter-
mined.
stituent
d-Galactose occurs most widely in the animal kingdom as a conIt has also been recognized by different of milk sugar.
investigators among the saponification products of protagon, a conThe galactose in protagon is supstituent of nerve and brain tissue. posed to be part of an amino-phosphoric-fatty acid complex, the exact
constitution of which
is
unknown.
d-Galactose has also been reported
to be present in different nucleo-proteids, mucins and other substances of animal origin, but the identity of the sugar in some of these cases has not been fully established.
The synthesis of d-galactose has Synthesis of d-Galactose. been accomplished in several ways. It has been built up by Fischer and Ruff* from d-lyxose, which, by addition of hydrocyanic acid and
saponifying (Kiliani's cyanhydrine synthesis), gives d-galactonic and d-talonic acids, the former, however, in much greater amount, f The lactone of d-galactonic acid upon reduction gives d-galactose.
also" been formed by Lobry de Bruyn and van Ekenby heating the ketose sugar 1-sorbose with dilute alkalies, a mutual rearrangement taking place between these two sugars similar to that noted between d-glucose and d-fructose. d-Galactose is Preparation of d-Galactose. From Milk Sugar. most easily prepared by hydrolyzing milk sugar. For this purpose
d-Galactose has
stein |
1 part of milk sugar is heated with 10 parts of 2 per cent sulphuric acid in a boiling water bath for 4 hours; the free acid is then neutralized with an excess of calcium or barium carbonate and the solution
trate
from the insoluble residue of sulphate and carbonate. The filevaporated to a sirup which will usually crystallize within a few days; crystallization may be hastened by priming the sirup with a The impure galactose crystal of galactose from a previous preparation.
filtered
is *
Ber., 33, 2142.
t Fischer calls attention to the fact
two acids
t
is
that in Kiliani's synthesis the yield of the never the same, one isomer being always produced in larger amount
1.
(Ann., 270, 64).
Rec. trav. Pays-Bas, 19,
THE MONOSACCHARIDES
from the
first
603
little 80 per hot water as possible; hot strong alcohol is then added, the solution boiled with a little bone black and filtered; the filtrate upon cooling will soon deposit crystals
crystallization
is filtered off,
washed with a
cent alcohol and then redissolved in as
little
of pure d-galactose. Preparation of d-Galactose
from Agar-agar.
d-Galactose
may
also
be prepared by hydrolysis of plant materials rich in galactan, as AgarThe latter when heated with 10 parts of 2 per cent sulphuric acid agar. for 12 hours in a boiling water bath is largely hydrolyzed to d-galactose which may be crystallized by neutralizing the acid solution and evaporating to a sirup as described in the preceding method. d-Galactose crystallizes from water Properties of d-Galactose.
as a monohydrate, CeHÔe.HÔ, in the form of large prismatic needles, and from strong ethyl and methyl alcohols as the anhydride in the form
of fine hexagonal crystals. the anhydride at about 165
The hydrate melts at 118 to 120 C. and C. The sugar has a sweet taste, is easily
soluble in water, moderately soluble in 50 per cent alcohol, but practically insoluble in absolute alcohol and ether.
d-Galactose
rotation
is
is
strongly dextrorotatory;
the
value
for
constant
about [a] D = + 81, the figure being influenced both by temThe sugar shows strong perature and concentration (see p. 181). mutarotation, [a] D immediately after solution being about +140. Tanret * has prepared d-galactose in a modification of low specific rotation. By dissolving 12 gms. of ordinary d-galactose in 30 gms. of water, adding 0.03 gm. of sodium phosphate exactly neutralized with sulphuric acid, heating a few minutes on the water-bath and then, after cooling, strongly agitating with 200 c.c. absolute alcohol, crystals of a galactose are obtained which give after solution a value for [a] D of only +53; this low rotation, however, increases upon standing of the solu-
The transition is tion to the true constant value of ordinary galactose. effected at once by heating the solution or by adding a trace of alkali
just as with the high rotating form of galactose. Action of Alkalies upon d-Galactose. By the action of dilute
alkalies d-galactose is
d-talose, d-tagatose, 1-sorbose several sugars.
transformed into a mixture of isomeric hexoses, and galtose as described under these
the action of 8 normal sodium hydroxide Nef f obtained 40 to 45 cent per d, 1-lactic acid, 10 per cent a- d-galacto-metasaccharin, 5 to 10 cent 0- d-galacto-metasaccharin, 5 per cent a-and /?- d-isosaccharin, per The two metasaccharins, which and numerous other saccharins.
By
Bull. soc. chim.
[3],
15, 195.
Ann. 376,
1.
604
are the
SUGAR ANALYSIS
most important
2
of the group,
have the following structural
formulae:
CH OH HOCH HC HCH
AJ. \_yjL A.
CH,OH
HOCH HC HCH
.LJ.V^JL
[OCH
HC( JOH
oift-
a- d-Galactometasaccharin.
d-Galacto-metasaccharin.
[a]g~-45.3
m. p.
144 C.
is
m.
[a]g=- 62.96. = 55 - 60 p.
C.
The
a- d-isosaccharin
described under lactose.
is fermented by yeast in presence and carbon dioxide; the fermentation proceeds, however, more slowly than with d-glucose and requires
Fermentation.
d-Galactose
of suitable nutrients to alcohol
about 8 days for completion. The yield of ethyl alcohol with cultures of pure yeast is about 45 to 46 per cent of the weight of galactose taken. * According to Buchner and Rapp the alcoholic fermentation of d-galactose is due to the enzyme zymase. Different species of Mucor and other moulds also cause alcoholic fermentation of d-galactose.f Many lactic acid producing organisms cause fermentation of d-galactose with formation of d, 1-lactic acid; with some organisms the 1-lactic acid is produced in greater amount. Bacterium xylinum, the so-called sorbose bacterium, oxidizes d-galactose to d-galactonic acid. Tests for d-Galactose.
Mucic Acid Reaction.
The
test
generally employed bined form, is the production of mucic acid

acid.
for detecting d-galactose, either in the free or
most comnitric
upon oxidation with
The
reaction
is
carried out as described under the determination
of galactan (p. 459). cent of its weight as
mucic
d-Galactose by this method t yields over 75 per acid. It must be remembered that mucic
1-galactose, dulcite
acid
is
also
formed by the oxidation of
and d- and
1-galactonic acids so that other reactions, such as the isolation of the sugar from its hydrazone, or the fermentation test, must be used for
confirmation.
* t
Ber., 31, 1090.
The idea of Dubrunfaut that shown by Pasteur to be erroneous.
d-galactose
later
was not fermented by yeast was view of Bourquelot that d-galactose
could be fermented only in presence of glucose, or some other easily fermentable sugar, was completely disproved by Tollens and Stone. J Tollens and Kent, Ann., 277, 222.
THE MONOSACCHARIDES
Mucic add has the configuration:
605
COOH HOCH HCOH HCOH HOCH ioOH

It crystallizes in
heating at 212 to 215 300 c.c. cold water).

structure,
is
minute granular rhombic prisms, which melt on rapid C. and are almost insoluble in water (1 part in Mucic acid, as is evident from its symmetrical
bromic
optically inactive. Upon heating with concentrated hydroacid, or other dehydrating agents, mucic acid is converted to
(p.
dehydromucic acid
781).
Reactions of d-Galactose with Phenylhydrazine. d-Galactose when treated with phenylhydrazine in the cold in the proportion of 5 parts sugar, 3 parts water and 5 parts phenylhydrazine deposits within
an hour a thick crystalline mass of d-galactose-phenylhydrazone * C 6 Hi 2 5 N 2 HC 6 H 5 After 24 hours the crystals are filtered off, washed with a little ether and recrystallized from hot alcohol; d-galactose phenylhydrazone forms fine colorless needles, melting at 158 C., easily soluble in hot water and alcohol, but insoluble in ether and chloroform.
: .
2 per cent aqueous solution is levorotatory f ([a] D = The 21.6). reaction be used for from phenylhydrazone may separating d-galactose d-glucose and other sugars whose hydrazones separate more slowly.
The
Methylphenylhydrazine and /3-naphthylhydrazine also form with d-galactose insoluble hydrazones which may be employed for purposes
of identification.
d-Galactose may be separated from its hydrazones by decomposing the latter with formaldehyde or benzaldehyde according to the usual
method.
d-Galactose, or its phenylhydrazone, upon heating with an excess of phenylhydrazine is converted into d-galactose-phenylosazone. | The latter forms fine yellow needles, having the formula CeHioO^^HCeHs^ and melting at about 194 to 196C.; presence of slight impurities
may
cause, however,
marked deviations
is
in the melting point (170
to
but slightly soluble in cold water; it is more soluble in hot water and is readily dissolved by hot 60 per cent
190 C.).
d-Galactosazone
alcohol.
*
f Jacobi,
Ann., 272, 171.
J Fischer, Ber., 17, 579.
606
SUGAR ANALYSIS
dulcite,
Reduction of d-galactose with sodium amalgam gives the alcohol which has the configuration
:
CH OH HOCH Hi OH HCOH HOCH CH OH

2 2
As
is
evident from
its
symmetrical
structure, dulcite
is
optically
inactive.
Dulcite
is
found in nature in Madagascar manna, Melampyrum
nemorosum and many other
Its melting point is 188 C. plants. Oxidation of d-galactose with bromine in aqueous solution gives d-galactonic acid, the lactone of which immediately after solution 70. shows a rotation of about [a] D =
1-Galactose.
CH OH HCOH HOCH HOCH HCOH HO

2
lactose
1-Galactose has been found in nature as a constituent of d, 1-gaamong the hydrolytic products of several plant materials; in
gum Chagual by
Winterstein* and in the Japanese food product, Nori, Tollens and Oshima.f by 1-Galactose has been prepared synthetically | by reducing the lactone of 1-galactonic acid which is formed together with d-galactonic acid by reduction of mucic acid with sodium amalgam. has been obtained as a white crystalline ^1-Galactose at 162 to 163 sugar melting C., easily soluble in water and 60 per cent alcohol, but only very slightly soluble in absolute alcohol. The
Properties.
sugar
is
is
strongly levorotatory,
[a] D
*
[a] D
74 about; strong mutarotation
observed,
8 minutes after solution
120.
Ber., 31, 1571.

t Fischer
t Ber., 34, 1422.
and Hertz,
Ber., 26, 1247.
THE MONOSACCHARIDES
1-Galactose
differs
is not fermented by yeast; from the behavior of d-galactose.
607
in this respect the sugar
This property renders
it
easy to detect 1-galactose in the presence of d-galactose, d-glucose, d-mannose, d-fructose and other fermentable sugars.
Tests.
1-Galactose
is
oxidized
by
strong nitric acid to
reduced by sodium amalgam to dulcite and is mucic acid, the sugar in both these re-
actions behaving the same as d-galactose; this agreement in behavior, in fact, follows necessarily from the configurations. 1-Galactose is oxidized with bromine to 1-galactonic acid, whose lactone (not isolated
as yet in the pure condition) is dextrorotatory. 1-Galactose forms with phenylhydrazine a difficultly soluble hydrazone melting at 158 to 160 C. and in appearance and solubility very
similar to d-galactose-hydrazone.
The aqueous
solution of 1-galactose
hydrazone,
guishes
it
sembles in
however, dextrorotatory, [o:] 21.6, which distinfrom d-galactose hydrazone; 1-galactose-phenylosazone reits appearance, melting point and solubility the osazone of
is
= D +
d-galactose.
Inactive galactose, as previously noted, has d, 1-Galactose. been found in a few cases among the hydrolytic products of certain
plant materials. The sugar has been prepared synthetically * by reducing the lactone of d, 1-galactonic acid, which is itself derived by reduction of the lactone
of
mucic acid
dulcite with
d, 1-galactose has also been prepared f by oxidizing hydrogen peroxide in presence of iron salts.
;
CH OH
2
CH OH
2
CHO
HOCH HCOH HCOH 2 +0 = HCOH HCOH HOCH HOCH CH OH CHO

I
HOCH
HOCH HCOH +2H -f HCOH HOCH CH OH

!
Dulcite
d-Galactose
1-Galactose
d, 1-Galactose, as obtained by Neuberg and Wohlwas found to have all the properties of a true racemic subgemuth, | stance. The sugar was obtained crystalline, melted at 143 to 144 C., was optically inactive and was fermented only one-half by yeast, the
Properties.
1-galactose remaining unattacked.

*
Fischer and Hertz, Ber., 25, 1247.
Neuberg and Wohlgemuth,
Z. physiol.
Chem.,
36, 219.
} Ibid.
608
SUGAR ANALYSIS
Tests. d, 1-Galactose is reduced by sodium amalgam to dulcite and oxidized by strong nitric acid to mucic acid, these reactions being, of course, the same as obtained by each of the component sugars. With phenylhydrazine d, 1-galactose forms an insoluble hydrazone, which separates rapidly from cold solutions of the sugar and, when
melting at 158 to 160 C. The hydrazone is decomposed by heating with formaldehyde or benzaldehyde with liberation of the free sugar, which may then be identified by its optical inactivity, by formation of mucic acid with nitric acid and by leaving a residue of 1-galactose after fermentation with yeast. Oxidation of d, 1-galactose with bromine gives d, 1-galactonic acid. The latter can be resolved into its components by fractional crystallizapurified, consists of colorless crystals
tion of its strychnine salt, the strychnine compound of d-galactonic acid separating as a crystalline deposit while that of 1-galactonic acid
remains in the mother liquor.
d-Gulose.
CH OH
2
HCOH HOCH HCOH HCOH CHO

d-Gulose has not been identified with certainty in any plant or animal product. The sugar has been prepared synthetically* by reduction of d-saccharic acid, which is converted first to the aldehyde compound
d-glucuronic acid, and then to d-gulonic acid.
CH OH HCOH HCOH HCOH HOCH + H = HOCH + H = HOCH + H O HCOH HCOH HCOH HCOH HCOH HCOH COOH COOH COOH
2
COOH
CHO
d-Saccharic acid
d-Glucuronic acid
d-Gulonic acid
The
lactone of d-gulonic acid
upon reduction with sodium amalgam
gives d-gulose.
*
Fischer and Piloty, Ber., 24, 521.
THE MONOSACCHARIDES
d-Gulose
is
609
upon d-sorbose
also
formed* by the action
of dilute alkalies
and
d-idose.
d-Gulose was obtained by van Ekenstein and Blanksmaf Properties = as white crystals melting at 165 C. and giving a rotation of [a] D
+ 42.9.
Tests.
The sugar
is
not fermentable.
d-Gulose gives upon reduction with sodium amalgam d-sorbite and upon oxidation with nitric acid d-saccharic acid. In these respects the sugar behaves the same as d-glucose, as follows necessarily from the configuration of the two sugars.
Oxidation of d-gulose with bromine in aqueous solution gives 55 d-gulonic acid, the lactone of which is dextrorotatory ([a] D =
about).
The phenylosazone
of d-gulose
is
identical with that of d-idose
and
of d-sorbose, these three sugars standing in the same structural relationship to one another as d-mannose, d-glucose and d-fructose.
1-Gulose.
CH OH
2
HOCH
HCO1 >H
HOCH HOCH CHO

1-Gulose has not been found as yet either free or combined in any natural product. The sugar has been prepared synthetically from 1-saccharic acid in the same manner as d-gulose from d-saccharic acid.
1-Gulose has also been built
up from
1-xylose
by the addition
of hydro-
cyanic acid and saponification of the nitrile; two acids are formed, as always, in this synthesis, 1-idonic and 1-gulonic. The lactone of the
latter
upon reduction gives

1-Gulose
1-gulose.f
Properties.
has been obtained only as a sweet levo-
= 20.4 about). rotatory unfermentable sirup ([a] D Tests. 1-Gulose gives upon reduction with sodium
bite
amalgam
1-sor-
and upon oxidation with nitric acid 1-saccharic acid. Oxidation with bromine in aqueous solution gives 1-gulonic acid, whose lactone consists of large rhombic hemihedral crystals melting at 181 C. and
showing levorotation
* t
([a] D
55 about).
Oxidation of the lactone

1.
Lobry de Bruyn and van Ekenstein, Rec. trav. Pays-Bas, 19, Rec. trav. Pays-Bas, 27, 1.
and Stahel,
Ber., 24, 528.
$ Fischer
610
SUGAR ANALYSIS
with hydrogen peroxide and basic ferric acetate gives 1-xylose, the starting point for the synthesis of 1-gulose. The phenylosazone of 1-gulose is identical with that of 1-idose and
of 1-sorbose.
*
of d, 1-gulonic of the lactones of d- and parts prepared by mixing equal acid, as been obtained a has The acid. only sirup. sugar 1-gulonic The lactone of d, 1-gulonic acid crystallizes in prisms melting at
d,
1-Gulose
is
is
obtained
by reducing the lactone
which
160 C. The crystals as ordinarily obtained from aqueous solution show opposite hemihedry f and the opposite forms when isolated belong Such crystals represent, of course, a mixture to the separate lactones. and not a true racemic combination, which should show only one
crystalline form.
(See page 785.)
d-Idose.
CH OH
2
HCOH HOCH HCOH HOCH
d-Idose has not been found in nature either in the free or combined
It has been prepared synthetically by Fischer { by reducing the lactone of d-idonic acid, which can be obtained through molecular rearrangement by heating d-gulonic acid with pyridine to 140 C. The
form.
sugar
is
also
formed by action of dilute
alkalies
upon d-gulose and
d-sorbose.
d-Idose has been obtained only as a clear Properties and Tests. non-fermentable sirup. Reduction with sodium amalgam gives the alcohol d-idite and oxidation with nitric acid gives d-idosaccharic acid,
which has been obtained only as a sirupy mixture of the acid and lactone ([a] D = over 100). d-Idose-phenylosazone is identical with that of d-gulose and d-sor-
bose.
*
t
Fischer and Curtiss, Ber., 25, 1025.
Haushofer, Ber., 24, 530; 25, 1027. t Fischer and Fay, Ber., 28, 1975.
THE MONOSACCHARIDES
1-Idose.
611
CH OH HOCH
2
HCOJ )H
HOCH HCOH CHO

1-Idose has not been discovered as yet in nature. The sugar has been prepared synthetically * by reducing the lactone of 1-idonic acid which is prepared from 1-xylose by addition of hydrocyanic acid and saponifying the nitrile (see under 1-gulose). 1-Idose has been obtained only as a colorless Properties and Tests. non-fermentable sirup ([a] D Reduction with sodium amal+7.5). gam gives the alcohol 1-idite and oxidation with nitric acid gives 1-idosaccharic acid, which has been obtained only as a sirupy mixture of acid and lactone ([a] Z) = over 100). 1-Idose-phenylosazone is identical with the phenylosazones of
1-gulose
and
1-sorbose.
d-Talose.
CH OH HOCH HCOH HCOH HCOH CHO

2
The sugar has been prepared

of d-talonic acid,
d-Talose has not been discovered as yet in any natural product. synthetically f by reducing the lactone
which can be obtained through molecular rearranged-Talose
ment by heating d-galactonic acid with pyridine to 150 C. is also formed by action of dilute alkalies upon d-galactose.
Properties
([a] D
and
.
Tests.
=+ 13.95)
idation with nitric
d-Talose has been obtained only as a sirup Reduction with sodium amalgam gives d-talite and oxThe latter forms microscopic acid d-talomucic acid.
29.4. crystals melting at 158 C. and showing dextrorotation, [a] D Oxidation of d-talose with bromine in aqueous solution gives d-talonic
*
Fischer and Fay, Ber., 28, 1975.
t Fischer, Ber., 24, 3622.
612
acid,
SUGAR ANALYSIS
which has been obtained only as a levorotatory sirup consisting of
is
the free acid and lactone.
d-Talose-phenylosazone
the same as that of d-galactose, this
identity following from their structural relationship.
1-Talose.
CH OH
2
HCOH HOCH HOCH HOCH CHO

1-Talose has not been found as yet in nature, nor has its synthesis been accomplished so far as known.
KETOHEXOSES
D-FRUCTOSE.
Levulose.
Fruit sugar.
2
CH OH HOCH HOCH
.HCOH
=O
H OH
2
one of the most abundant and widely In the free condition it is almost distributed sugars found in nature. always associated with glucose as a constituent of plant juices, such as the must of fruits, the sap of green leaves and stalks, and the nectar of flowers. Owing to the fact that d-glucose and d-fructose occur so often in very nearly equal amounts, it is supposed that the two sugars
Occurrence.
d-Fructose
is
are largely formed
of inverting enzymes upon sucrose. fructose to glucose and sucrose in the mixed The relationship of sugars of different plant juices may be seen from the following table:
by the action
'THE
MONOSACCHARIDES
613
d-Fructose is also widely distributed in nature as a constituent of various anhydride condensation products, such as complex sugars and
polysaccharides. Among the complex sugars, which give d-fructose upon hydrolysis with acids or enzymes, may be mentioned sucrose, raffinose, lupeose, stachyose, secalose and gentianose.
In addition to the complex sugars there are a large number of

plant constituents of a
gummy character which give d-fructose upon These substances, which occur mostly as a reserve mahydrolysis. terial in the tubers and root organs of several families of plants, are
sometimes called inuloids from their chief representative inulin. The inuloids have the same general formula (C 6 Hi 05) n with varying amounts of water of hydration and are levorotatory, the values for 20 to [a] D ranging from about 50; they are all soluble in hot water, from which solution they are precipitated by absolute alcohol or by the hydroxides of the alkaline earths. The inuloids obtained from different sources are no doubt in very many cases identical, the differences in analysis, specific rotation, melting point, etc., being probably due
to
accompanying impurities.
Inulin.
Inulin occurs very widely distributed as a reserve
ma-
the root organs of the Compositse and allied families of plants such as the Campanulacese, Lobeliaceae, etc. It was discovered by
terial in
Rose* in 1804 in the roots of Inula Helenium (elecampane), from which plant the compound derives its name. The tuberous roots of the dahlia, dandelion, chicory, Jerusalem artichoke (Helianihus tuberosus], arnica and pyrethrum are other examples of plant materials rich in inulin. Owing to its use by plants as a reserve material the percentage of inulin in roots and tubers is subject to wide fluctuations, being usually least in spring and greatest in autumn. Dandelion roots,
for example,
in
were found by Dragendorff f to contain 1.74 per cent inulin
March and 24
ethrum and Jerusalem artichoke

in the
The dahlia, chicory, pyrper cent in October. may contain over 50 per cent inulin
dry substance of the roots.
Inulin may be prepared according to Preparation of Inulin. Kiliani % by reducing to a fine pulp the ripe tubers (taken in autumn) of the dahlia, chicory, etc., boiling the material with water in presence of calcium carbonate and filtering. The filtrate is then frozen in a freezing mixture
and
*
t t
after
thawing out the precipitated inulin
filtered off;
"
Gehlen's Neues allgem. J. Chem., 3, 217. " (1870). Monographic des Inulins
Ann., 206, 147.
614
SUGAR ANALYSIS
the raw product thus obtained is redissolved in hot water and again frozen out. After repeating the purification in this way for several times the final product is washed with 93 per cent alcohol, then with
little
ether and afterwards carefully dried in a water oven. Tanret* recommends a preliminary clarification of the hot root ex-
after filtering, the solution is freed from the inulin precipitated by adding a then lead by sulphuric acid and barium concentrated solution of hydroxide and heating. The precipitate is washed with cold barium hydroxide solution and then decomposed in aqueous suspension with carbon dioxide; the solution after heating is filtered from barium carbonate, and the inulin precipitated from the tracts with lead subacetate;
filtrate
by means
of strong alcohol.
Fig. 194.
Sphere-crystals of inulin (precipitated from cells of the dahlia

of alcohol)
.
by means
Properties of Inulin.
position
of
tiple thereof.
Inulin, as generally prepared, has a
com-
the formula 6(C 6 Hio0 5 )
+ H O,
2
i.e.,
CseHÔsi or a mul-
Brown and Morris f
give the formula C72Hi 2 40 6 2 and
Inulin consists of a white hygroscopic substance, Oi55. soluble in hot water, in which it may form supersaturated very easily solutions. It is insoluble in absolute alcohol. Its aqueous solution
Tanret Ci 8oH 3 i
does not reduce Fehling's solution, and gives no color reaction with iodine (distinction from soluble starch, plant glycogen, and dextrin). = 38 (about), the Inulin in aqueous solution is levorotatory, [a]
values as determined
by
ranging from
*
36 to
different authorities for different preparations 40.

201.
f
Bull. soc. chim.
[3], 9,
Chem. News,
69, 296.
THE MONOSACCHARIDES
615
Inulin is rapidly hydrolyzed (15 to 20 minutes) to d-fructose upon heating with dilute acids. Hydrolysis may also be effected by heating with water alone under pressure at 110 to 120 C. A special enzyme
inulase which
is
found in the germinating tubers of the Jerusalem
arti-
choke, and other inulin-containing plants, also hydrolyzes inulin to d-fructose; other enzymes such as diastase, invertase, emulsin, etc., Inulin is not fermented by yeast. are without action.
Inulin can usually be detected in plant tissues
by placing thin
sections of the tubers, etc., in strong alcohol or glycerol and then examining the preparation under the microscope. The inulin will be
precipitated within the cells as sphere-crystals

fissures (Fig. 194).
marked with
radial
In addition to inulin Tanret,* by the fractional precipitation of the extract from Jerusalem artichokes with barium hydroxide, has separated the following closely allied compounds:
Pseudoinulin Inulenin Helianthenin
32.2 29.6 23.5

17.0.
Synanthrin
terials
Among may
other inuloid substances f found in different plant be mentioned the following:
ma-
Compound.
616
SUGAR ANALYSIS
commonly than d-glucose, and frequently only as a result of disease It occurs at times in normal urine, as or other abnormal condition.
meats or drinking sweet wines, found in the urine of certain diabetic patients; such urine even when rich in fructose may show but little levorotation owing to the counter effect of the rotation of d-gluLevorotation in urine may also be produced by glucuronic acid cose. complexes (p. 375), so that an optical examination of urine without confirmatory tests is not always to be relied upon. The occurrence of d-fructose in honey Honey and Floral Nectar. has already been referred to. The average amount of fructose, glucose and sucrose in honey according to different authorities is given as folafter eating excessive
amounts
of sweet
is
champagnes,
etc.
d-Fructose
also
lows:
THE MONOSACCHARIDES
dilute alkaline solution.
logically
617
The
from d-mannite, which
synthesis can also be accomplished biois oxidized by the sorbose bacterium
almost quantitatively to d-fructose.*
CH OH
2
HOCH HOCH HCOH * HCOH CH OH

|
CH OH HOCH HOCH +O = +H HCOH

2
I
C=O
CH OH
2
d-Mannite
d-Fructose
The oxidation
place
(p.
of d-mannite
771) in the second or
fifth position
by the sorbose bacterium may take (marked by a*) with for-
mation of d-fructose in either
case.
d-Fructose is most easily prepared Preparation of d-Fructose. by hydrolyzing some one of its condensation products. For this purpose sucrose and inulin are the substances generally chosen. The sugar
on account of
its extreme solubility is very difficult to crystallize. In the preparation f from Preparation of d-Fructose from Sucrose. sucrose a solution of invert sugar is prepared by inverting a 10 per cent solution of sucrose at 60 C., using for every 100 gms. of sucrose
c.c.
about
of concentrated hydrochloric acid. The solution after cooling to 5 C. is treated for each 100 c.c. with 6 gms. of fresh finely
After stirring the solution vigorpulverized pure calcium hydroxide. ously for 2 to 3 minutes, the liquid is filtered rapidly using a cooling
funnel
the filtrate which should be kept cold soon deposits fine needles of calcium fructosate. The latter after standing 24 hours is filtered off,
;
using a centrifuge or Buchner funnel, washed with ice water and then after suspending in water at 20 C. carefully decomposed with the exact amount of oxalic acid. Any excess of the latter may be removed by addition of a little of the calcium fructosate kept back as a reserve.
The
from the calcium oxalate is then evaporated at low temThe latter after priming with a a vacuum to a sirup. of fructose from a previous preparation and setting aside in crystal
filtrate
perature in
a cool place over concentrated sulphuric acid will usually crystallize within a few days. Crystallization may also be effected by dissolving as much as possible of the concentrated sirup in warm absolute
*
t
Brown,
J.
Chem.
Soc., 49. 172.
Modification of the original method of Dubrunfaut, Compt. rend., 25, 307;
69, 1366.
618
alcohol,
SUGAR ANALYSIS
and then pouring
off, when cold, the soluble portion from the solution upon standing will soon deposit alcoholic the sirupy residue;
crystals of pure d-fructose.
d-Fructose is most easily this 100 For inulin. from gms. of inulin and 250 purpose* prepared c.c. of water are heated with a little hydrochloric acid for 30 minutes in a boiling water bath. The quantity of hydrochloric acid used for hydroPreparation of d-Fructose from Inulin.
lyzing depends upon the ash content of the inulin; for 100 gms. inulin of 1 per cent ash content 0.5 gm. HC1 is taken, for 0.2 per cent ash 0.1 gm. HC1 and if the inulin is ash free 0.01 gm. HC1. After hydrolysis the
free acid
is
neutralized with an excess of calcium carbonate and the
solution filtered.
The
filtrate is
is
perature to a thin sirup, which
evaporated in a vacuum at low temthen set aside in a vacuum desiccator
over concentrated sulphuric acid. After standing some days the thick sirup is warmed with absolute alcohol and after thorough agitation
allowed to stand 24 hours. The clear alcoholic solution is then poured off, primed with a few crystals of fructose and set aside in a cool place.
Crystallization perfectly white black.
is
by
usually complete in 3 days. The sugar is obtained recrystallizing from hot absolute alcohol using bone
d-Fructose crystallizes from absolute Properties of d-Fructose. ethyl or methyl alcohol as the anhydride C 6 Hi 2 6 in fine colorless A crystalline hydrate of the formula needles melting at 95 to 105 C.
exceedingly soluble in cold water, but only very slightly soluble in cold absolute alcohol. It is easily soluble in hot absolute
is
(C 6 Hi 2 O 6 )2 d-Fructose
+H
has also been obtained.
Unlike most sugars d-fructose ethyl and methyl alcohol. a considerable extent in mixtures of alcohol and ether.
d-Fructose
is
is
soluble to
92 although very strongly levorotatory, []^ = changes in temperature and concentration may produce considerable
variations from this figure as shown on page 181. Diluting a concentrated fructose solution with water causes a low-
ering of the specific rotation, and about 30 minutes are necessary for the reading to become constant.
d-Fructose exhibits mutarotation, the value for
[a] D
immediately
after solution being about The change to constant rotation pro106. ceeds much faster than with other mutarotating sugars, and is usually
completed within an hour.
The
effect of acid in increasing the levorotation of d-fructose

to.
*
has
been referred
Ost. Z. analyt.
Chem.,
29. 648.
THE MONOSACCHARIDES
Fermentation of d-Fructose.
d-Fructose
is
619
fermented in the
same manner
by various yeasts, moulds and bacteria. Yeast produces about the same yield of alcohol and carbon dioxide
as d-glucose
from d-fructose as from d-glucose, but the fermentation in
its
first
The alcoholic fermentation stages proceeds more rapidly with glucose. of fructose by means of the enzyme zymase has been accomplished by
Buchner
in the
same manner
as for glucose.
d-Fructose undergoes the lactic and butyric fermentations with

the same readiness as d-glucose. In certain anaerobic fermentations where free hydrogen is evolved d-fructose is reduced to d-mannite, the reaction being of the same
amalgam and other reducing mannite by microorganisms in fructoseagents. mannitic is often termed a fermentation. solutions containing Tests for d-Fructose. d-Fructose upon reduction with sodium amalgam yields equal parts of d-mannite and d-sorbite.
The formation
of
character as that obtained with sodium
CH OH
2
620
SUGAR ANALYSIS
are heated to a high temperature the with formation of oxymethylfurfural. partly decomposed
If solutions of d-fructose
sugar
is
COH
C H O8 =
6
12
HO
2
d-Fructose
Oxymethylfurfural.
The oxymethylfurfural formed Tests for Artificial Invert Sugar. in the previous reaction is easily detected by its coloring aniline acetate red or
by its forming brilliant colorations with phloroglucin, resorcin and other phenols. This property has been made use of for detecting artificial invert sugar in honey, and other food products. Artificial
is
invert sugar
solutions with a small
0.1 per cent of weight of sucrose) to 110 to 120 C., at which temperature perceptible amounts of oxymethylfurfural are formed.
made commercially by heating concentrated amount of tartaric or other acid (about
sucrose
In making the test for oxymethylfurfural Fiehe* rubs up the product (honey, etc.) with ether and filters the ethereal solution into
a small porcelain dish. After evaporating the ether, the residue is heated with a 1 per cent solution of resorcin in concentrated hydrochloric acid. In presence of artificial invert sugar a red color develops which soon changes to a reddish brown. A more rapid but less sensitive reaction for artificial invert sugar is obtained with aniline acetate. f The reagent, which should be
freshly prepared before using, is made by shaking up 5 c.c. of chemically pure aniline with 5 c.c. of water and adding sufficient glacial acetic acid (2 c.c.) to just clear the emulsion. In making the test 5 c.c. of a concentrated solution of the honey, etc., are treated in a test tube with
1 to 2 c.c. of the aniline reagent. The latter is allowed to flow down the walls of the tube so as to form a layer upon the surface of the solu-
tion underneath.
If
a red ring forms beneath the aniline solution,
when the tube
gently agitated, oxymethylfurfural is present. It should be borne in mind that honeys or other fructose-contain-
is
ing products which have been cooked or boiled also give the reaction
for oxymethylfurfural.
The
brilliant red coloration
obtained upon heating d-fructose or
sucrose with concentrated hydrochloric acid

*
and sesame
Chem.,
oil is
probably
Z. Nahr. Genussmittel, 16, 75.
Browne,
Bull., 110,
U.
S. Bur.
p. 68.
THE MONOSACCHARIDES
constituent of the
oil.
621
due to a condensation product between oxymethylfurfural and some
Hydrobromic
acid reacts with fructose in ether solution to form
C 4 2 O CHO, which colors the solution 2 Br bromomethylfurfural and reddish a deep crystallizes in gold colored prisms. purple (Reaction of Fenton and Gostling.*) This reaction is also given by other
sugars and carbohydrates, but is most pronounced with those which contain a fructose group. d-Fructose is more sensitive in Reducing Reactions of d-Fructose.
CH
reducing power than most other sugars and this property has been utilized as a means of identification.
Pinoff f
recommends
for the
above purpose a 4 per cent solution

of the latter diluted with acetic acid gives
of
ammonium molybdate;
water containing 0.2
0.1
10
c.c.
10
c.c.
of
c.c. of glacial
upon heating with
gm. d-fructose in a water bath at 95 to 98 C. a bright blue coloration; solutions of other sugars under these conditions remain colorless. Any free mineral acid must be neutralized before conducting the exgive the reaction. a solution of copper in in carbonate or alkaline amino-acetic acid hydroxide potassium is The latter reagent (glycocoll). prepared by dissolving 12 gms. of in 6 of glycocoll water; gms. copper hydroxide are then added gradu-
periment, otherwise other sugars

Pieraerts {
may
recommends
for detecting fructose
ally
and when solution is complete the hot liquid is cooled to 60 C.; 50 gms. of potassium carbonate are then added and the solution made up to 1 liter. In testing for fructose the product to be examined is
dissolved in cold water, clarified if necessary with a little lead acetate, filtrate freed from excess of lead by means of sodium sulphate and the clear solution diluted to about 5 per cent reducing sugar. Upon
will
the
heating with the alkaline glycocoll-copper solution to 30 C. reduction take place within an hour if fructose is present; reduction is also obtained at ordinary temperature after 12 hours' standing. Other sugars are said not to show reduction under these conditions.
d-Fructose forms a Methylphenylosazone Reaction of d-Fructose. with number osazones of and phenylhydrazine and large hydrazones
its
substituted derivatives.
fication the osazone reaction
Neuberg
*
J.
For purposes of separation and identiwith methylphenylhydrazine is stated by to be of great value. d-Fructose-methylphenylosazone is
Soc., 73, 556; 75, 423; 79, 361.
t Ber., 38, 3317.
Chem.
J Belg.
Ann. de Pharmacie, March and
April, 1908.
Bull, assoc. chim. sucr. dist.,
25, 830.
Ber., 35, 959.
Z. physiol. Chem., 36, 227.
622
SUGAR ANALYSIS
in alcoholic acetic acid for a
obtained upon heating fructose solutions with methylphenylhydrazine few minutes and then setting aside in
Crystallization
is
a cool place.
almost complete within a few hours.
The compound consists of yellowish crystals having the composition C 2 oH2 6N 4 04, and melting between 158 and 160 C.; it is only slightly soluble in water, cold alcohol and ether, but is easily soluble in hot
alcohol, acetone
and chloroform.
Glucose, mannose, galactose and other aldose sugars do not form osazones with methylphenylhydrazine owing to the fact that the
CHO radicle is prevented in group adjoining the terminal some way from reacting with substituted hydrazines. In the case of CH 2 OH group occufructose and other ketoses, where the reacting pies the end position, the freedom of osazone formation is not impeded. d-Fructose-phenylosazone is identical with that of d-glucose and d-mannose. The formation of osazone is more complete, however, with d-fructose than with its aldose isomers (see page 350). d-Fructose reacts with alkalies similarly to d-glucose.
1-Fructose.
CHOH
CH OH
2
HCOH HCOH HOCH
|
;
A-O
CH OH
2
The sugar has been prepared synthetically* by reduction of 1-glucosone (obtained from 1-glucose-osazone), in the same manner as d-fructose is prepared from d-glucose-osazone 1-fructose can also be prepared from d, 1-fruc1-Fructose has not been found as yet in nature.
tose
yeast. 1-Fructose has been obtained only as a dextrorotatory unfermentable sirup. The sugar has not been separated in a pure crystallized
its
by fermenting away the d-component with
form, so that other knowledge of

ties is lacking.
chemical and physical proper-
d, 1-Fructose.
Inactive, or racemic, fructose has not been found
The sugar has been prepared synthetically, however, by a number of methods and some of these have a special interest in that the sugar was built up from simple organic compounds jiot belonging
in nature.
*
Fischer, Ber. 23, 373, 2618, 3889; 24, 2683.
THE MONOSACCHARIDES
623
The best known example of such a method is the classic to the sugars. * by which acrolein dibromide in consynthesis of Fischer and Tafel
barium hydroxide is condensed to form a-acrose and other hexose sugars. 2 Ba(OH) 2 = 2 BaBr2 C 6 12 6 2 C 3 H4Br2
tact with
Acroleindibromide
a-Acrose.
The above reaction is carried out in ice-water. The solution, after precipitating barium, is evaporated and treated with phenylhydrazine. Two osazones are formed, one insoluble in ether (a-acrose-osazone)
and the other soluble
osazone
is
in
ether
(/3-acrose-osazone).
The
a-acroseis
found to be identical with that of
d, 1-glucose,
which
also
the same as that of d, 1-mannose, or d, 1-fructose. a-Acrose-osazone upon treatment with zinc dust and acetic acid is reduced to a-acrose-amine, and the latter upon treatment with nitrous
acid
is
converted to d, 1-fructose.
CH OH
2
CH OH
2
(CHOH) 3
2
C=0 H C-NH
2
+2HN0 =2
2
(CHOH)s
C=0 CH OH
|
+2N + 2H
2
a-Acrose-amine
d.l-Fructoae.
less optically inactive sirup, easily soluble in
has been obtained only as a sweet colorwater and alcohol, but inIt reduces Fehling's solution and gives the other soluble in ether. common reactions of a reducing sugar. It is fermented only one-half with yeast, the 1-fructose remaining in an unaltered condition.
Properties.
d, 1-Fructose
Reduction of d, 1-fructose with sodium amalgam gives d, 1-mannite, the oxidation of which to d, 1-mannonic acid has been mentioned in the synthesis of d-mannose and d-glucose.
D-SORBOSE.
d-Sorbinose.
CH OH
2
HCOH HOCH
HCOI
Occurrence. Although d-sorbose has not been found free in the natural juices of plants this sugar has been discovered in large quan*
Ber., 20, 2566; 22, 97.
624
SUGAR ANALYSIS
titles in the fermented juice of sorb-apples (the fruit of the service-tree, Sorbus domestica) and other fruits of the rosaceous family. The sugar was discovered first by Pelouze,* but Boussingault f was the first to show that the sugar was formed by an oxidizing fermentation of the
hexite alcohol d-sorbite which
found in sorb-apples, mountain-ash the principal organisms concerned berries and other fruits. so-called is the sorbose bacterium (Bacterium in this fermentation which d-sorbite of is represented as follows: upon xylinwri), the action
is
One
of
CH OH
2
CH OH
2
HCOH HOCH HCOH * HCOH CH OH

I
+0=
HCOH HOCH +H HCOH C=0 CH OH

I
d-Sorbite
d-Sorbose
The oxidation takes place only
in the position
marked with a
*,
as
explained on page 771. d-Sorbose is formed synthetically in small amounts

of dilute alkalies
by the action
upon d-gulose and
d-idose.
d-Sorbose is prepared according to Preparation of d-Sorbose. Bertrand's J method by evaporating the juice of sorb-apples to a specific
by fermentation with
gravity of 1.05 to 1.06 and then removing all fermentable sugars When the alcoholic fermentation is comyeast.
pleted the clear solution is poured into shallow dishes, inoculated with a strong pure culture of Bacterium xylinum and allowed to stand at 30 C. until the reducing power of the solution has reached its maximum. The liquid is then clarified with lead subacetate, the excess of lead removed from the filtrate with the exact amount of sulphuric acid and the filtrate, which must be perfectly neutral, evaporated in a
vacuum
alcohol
to a sirup.
The
latter is purified in the usual
way by
is
strong
about 80 per cent of the d-sorbite originally present. Instead of using sorb-apple juice a solution of d-sorbite in presence of yeast extract, asparagine, peptone and mineral nutrients may be used for fermentation. By this method Lobry de Bruyn and van Ekenstein obtained a yield of about 30 per cent. d-Sorbose has been obtained in the form of colorless Properties. rhombic crystals melting at 154 C. The sugar is very sweet, easily
for crystallization.
and set aside
The yield of
d-sorbose
Compt. Compt.
rend., 34, 377.

rend., 74, 939.
Compt.
rend., 122, 900.
Rec. trav. Pays-Bas, 19, 3
THE MONOSACCHARIDES
625
soluble in water, but difficultly soluble in alcohol. d-Sorbose is levo= 42.5, this value being slightly influenced by changes rotatory, [a] D in temperature and concentration as shown on page 181.
d-Sorbose
yeast.
is
The sugar
not fermented by any of the ordinary varieties of is also very resistant to the attacks of moulds and
bacteria; certain lactic acid organisms, however, were found thelot * to produce lactic and butyric acid.
by Ber-
d-Sorbose is reduced by sodium amalgam to the alcohols and d-idite. Oxidation with nitric acid produces d-tartaric, oxalic and other acids. d-Sorbose reduces Fehling's solution and gives the characteristic color reaction of the ketoses with resorcin and hydrochloric acid. Upon heating with 3 parts of phenylhydrazine chloride and 5 parts of sodium acetate d-sorbose gives an osazone, CisH^N^, which is identical with that of d-gulose and d-idose. The osazone consists of
Tests.
d-sorbite
fine
yellow needles melting at 164 C.

1-Sorbose.
1-Sorbinose.
CH OH
2
HOC!
HCO] )H
HOCH i-o CH OH
2
Synthesis.
in the
1-Sorbose has not been found in nature either free or
combined form. The sugar has been prepared synthetically by Lobry de Bruyn and van Ekensteinf by warming d-galactose in 20 per cent aqueous solution with not more than 3 per cent potassium hydroxide for 3 hours at 70 C. The solution, which has acquired a weak acid reaction, is cooled and the unchanged galactose allowed to The mother liquor is then evaporated, and extracted with crystallize. methyl alcohol and acetone. The residue is then fermented to remove the rest of the galactose and the solution evaporated to a sirup, from which the 1-sorbose crystallizes after long standing. The yield is 6 to
8 per cent of the d-galactose taken. The above reaction by which 1-sorbose
of secondary rearrangements
is formed belongs to a class which are peculiar to many of the sugars. As d-glucose upon warming with dilute alkalies undergoes partial *
Ann. chim. phys.
[3],
60, 350.
t Rec. trav. Pays-Bas, 16, 262;
19, 1.
626
SUGAR ANALYSIS
rearrangement into d-mannose and the ketone sugar d-fructose, so d-galactose is transformed into d-talose and the ketone sugar d-tagaThe ketone sugars which are formed in these reactions seem, tose. however, on prolonged warming with alkalies to be partially trans-
formed into isomeric ketoses. The reaction between d-galactose, d-tagatose and 1-sorbose would be represented as follows:
CH OH
2
THE MONOSACCHARIDES
Tests.
627
1-Sorbose
is
alcohols 1-sorbite
and
1-idite.
reduced by sodium amalgam to the hexite The sugar reduces Fehling's solution
somewhat stronger than d-galactose and gives the characteristic color reaction of ketoses with resorcin and hydrochloric acid. The phenylosazone is identical with that of 1-idose and 1-gulose.
This sugar was prepared by Lobry de Bruyn and d, 1-Sorbose. van Ekenstein* by evaporating a solution containing equal parts of d- and 1-sorbose. The sugar was obtained in white crystals melting at
of its
A study by Adriani f of its solubility as compared with that two components showed that the crystals were a true racemic combination and not a simple mixture.
154 C.
d-Tagatose.
CH OH HOCH HCOH
2
HCOH
!=0
H OH
2
d-Tagatose has not been found as yet in nature either any combined form. The sugar has been prepared synthetically by Lobry de Bruyn and van Ekenstein I by the action of The dilute alkalies upon d-galactose as described under 1-sorbose.
Synthesis.
free or in
mother liquor
after crystallization of 1-sorbose yields upon evaporation a mixture of crystals consisting of 1-sorbose and d-tagatose. The mixed crystals are dissolved in 5 parts of absolute methyl alcohol and 2 parts aniline, from which the 1-sorbose crystallizes at once and the
d-tagatose after evaporating the mother liquor.
The sugar
is
purified
by
recrystallization.
Properties.
d-Tagatose
consists
of
white
is
124 C.
The sugar has a sweet
taste
and
crystals melting at easily soluble in water,
but
difficultly soluble in alcohol.
The aqueous
is
solution
is
very weakly
is
dextrorotatory, [a]g 2.6. rotatory, [a]^ =
= -f
at higher temperatures the sugar
levo-
not fermented by yeast. d-Tagatose with sodium amalgam Tests. reduction d-Tagatose gives upon Oxidation with nitric acid the hexite alcohols dulcite and d-talite.
*
t
Rec. trav. Pays-Bas, 19, 1. Rec. trav. Pays-Bas, 19, 185. t Rec. trav. Pays-Bas, 16, 62, 282; 18, 72.
628
SUGAR ANALYSIS
causes a disintegration of the carbon chain with formation of 1-tarThe sugar reduces Fehling's solution taric, oxalic and other acids.
somewhat stronger than d-galactose and gives the characteristic color reaction of ketoses with resorcin and hydrochloric acid. The phenylosazone is identical with that of d-galactose and d-talose.
1-Tagatose.
CH OH
2
HCOH HOCH HOCH i-o CH OH

2
The sugar is 1-Tagatose has not been found as yet in nature. formed by molecular rearrangement according to Lobry de Bruyn
and van Ekenstein* by the action of
d-tagatose.
dilute alkalies
upon
d-sorbose,
the transformation being the same as that between 1-sorbose and
The sugar has not been

properties
isolated as yet in the crystalline

Its
form and
is
its
are
therefore
with that of 1-galactose
unknown. and 1-talose.
phenylosazone
identical
d, 1-Tagatose. Racemic, or inactive, tagatose has not been found in nature, nor has the sugar up to the present been prepared An inactive ketose sugar f has been detected among synthetically. the oxidation products of dulcite obtained by action of lead peroxide,
and
this sugar in all probability consists in part at least of d, 1-tagatose.
KETOHEXOSES OF UNKNOWN STRUCTURE

Galtose. J
1-sorbose as being
This ketohexose has already been referred to under formed by the action of alkalies upon d-galactose
through secondary rearrangement.
The sugar remains
in the
mother
liquors after crystallization of the 1-sorbose
and d-tagatose.
tions
Galtose has been obtained only as a sweet sirup. Its aqueous soluhave but little rotatory power and are not fermentable. Galtose reduces Fehling's solution only about half as strong as d-galactose.
Distillation with hydrochloric acid gives 4 to 5 per cent of furfural.
t
Rec. trav. Pays-Bas, 16, 62, 282; 18, 72. Neuberg, Ber., 35, 2629. $ Lobry de Bruyn and van Ekenstein, Rec. trav. Pays-Bas, 16, 257, 262.
THE MONOSACCHARIDES
at 182
629
Galtose-phenylosazone, Ci 8 C.
H 2N
2
4 04,
forms yellow crystals melting
This ketohexose is formed by secondary rearrangeaction of dilute alkalies upon d-glucose, d-mannose the ment through and d-fructose. The best yields are obtained by heating a 20 per cent solution of d-glucose or d-fructose with 10 per cent of pure moist lead
Glutose.*
hydroxide for 3 hours at 70 C. The lead is then precipitated, the mixture of sugars fermented with yeast, when the glutose remains beThe yield of glutose by this method is about 20 per cent from hind.
d-glucose and 40 per cent from d-fructose. Occurrence of Glutose in Cane Molasses.
While glutose does not for in commercial be looked can always presence been d-fructose have and where subjected to the d-glucose products action of alkalies. It has been found in sugar-cane molassesf in amounts varying from 1 to 5 per cent as a result of the action of the lime used in clarification upon the invert sugar of the juice. Glutose, not being from molasses of the vinasse constituent is found as a fermentable,
occur in nature
its
distilleries.
of glutose
In the valuation of molasses for distilleries the amount and other non-fermentable reducing sugars should be detercarefully conducted fermentation test.
mined by a
Glutose has not been obtained in the crystalline Properties. It reduces so form, nothing is known of its physical properties.
Fehling's solution about half as strong as d-glucose and gives the characteristic color reaction of ketoses with resorcin and hydrochloric acid.
Aqueous solutions
of glutose
show no perceptible
optical activity.
Pseudofructose.
the products obtained
d-fructose.
This ketohexose has also been detected among
by action of dilute alkalies upon d-glucose or The sugar was obtained by Lobry de Bruyn and van
([a] D
Ekenstein t as a levorotatory sirup

isolated in the pure condition.
40 about) but has not been
Formose.
The condensation
of
alkalies to a sweet sugar-like substance
formaldehyde in presence of was first observed by Butlerow,
who gave
*
the
compound the name methylenitan.

||
similar condensait
tion product
was afterwards prepared by Loew

dist., 16,
who gave
the
name
Lobry de Bruyn and van Ekenstein, Rec.

chim. sucr. Rec. trav. Pays-Bas, 16, 162.
rend., 53, 143.
J.
trav. Pays-Bas, 16, 62, 282.
t Pellet, Bull, assoc. t
1181; 19, 834.
Compt.
II
prakt.
Chem.
[2],
33, 321.
630
formose.
SUGAR ANALYSIS
Methylenitan and formose are according to Fischer identical been disputed by Loew and Tollens. To 4 cent a solution of formaldehyde is shaken with formose per prepare an excess of milk of lime for half an hour and then filtered; the alkaline filtrate is allowed to stand for 5 to 6 days at room temperature when the condensation is complete. The solution is then neutralized with
in nature, although this has
and evaporated to a thin sirup; the latter is puriand other impurities by means of strong alcohol; the alcoholic solution upon evaporation gives a residue which conoxalic acid, filtered
fied
from lime
salts
sists
mostly of formose.
Formose has been obtained only as a yellowish insweet It tensely sirup; it is optically inactive and strongly reducing. is not fermented by yeast, although certain organisms decompose the sugar with formation of lactic acid. The sugar, from the analysis of
Properties.
its
resorcin
osazone, belongs to the hexoses, and from its color reaction with and hydrochloric acid is a ketose. The configuration of foris
it is still a question whether not a mixture * of sugars rather than a single substance. Formose-phenylosazone, C^H^N^, was obtained by Fischer f as fine yellow needles melting at 144 C.
mose has not as yet been determined and
formose
p-Formose { and morf ose are two other sugars which Loew has obtained from formaldehyde by varying the temperature and other conditions of condensation. The sugars have been obtained only as
impure sirups; the existence of these sugars has been strongly questioned.
Lycerose was obtained by Loew as an impure sirup through condensation of glycerose with calcium hydroxide at 75 C. In all probability lycerose is a mixture of several condensation products.
||
NATURAL HEXOSES OF UNCERTAIN CHARACTER
large
number
of hexose sugars of
unknown
literature.
been reported at various times in the
configuration have The existence of

possible that some of the hexoses
these in nearly all cases requires confirmation. of the sugars in the following list belong to
*
It
is
some one
is
According to Nef (Ann., 376,
1)
synthetic formose
possible aldo-
and keto-tetroses, -pentoses and -hexoses,
in the equilibrium
probably a mixture of all between
which some 116 different substances take part.

t Ber., 21, 988.
t J. prakt.
Chem.
[2],
34, 51.
II
Chem. Chem.
Ztg., 23, 542. Ztg., 23, 542.
THE MONOSACCHARIDES
631
previously described, the variations noted in specific rotation and other properties being due to impurities.
Sugar.*
632
SUGAR ANALYSIS
a-Rhamnohexoseserves to establish the configuration of the sugar. phenylosazone crystallizes in fine yellow needles insoluble in water, but
soluble in hot alcohol;
it
melts at 200 C.
p-Rhamnohexose.*
CH
:HOH
HC< ;OH
HOCH
HOC!
HOC]
a-Rhamnohexonic acid upon heating with pyridine to 150 to 155C. undergoes partial transformation to /3-rhamnohexonic acid whose lactone is reduced by sodium amalgam to /3-rhamnohexose. The sugar has not been isolated and its properties are for the most part unknown.
Oxidation with nitric acid splits
off
the methyl group with formation
of talomucic acid, this reaction serving to confirm the configuration. The phenylosazone of /3-rhamnohexose melts at 200 C. and in all other
respects
is
identical with that of a-rhamnohexose.
a-Rhodeohexose.j
CH
OH Hi OH
HCOH HOCH HCOH iHO

This sugar has been prepared from rhodeose in the same way that a-rhamnohexose was built up from rhamnose. The sugar consists of
fine crystals
melting at 125 to 126 C. and showing in aqueous solution

consists of golden needles melting at 231
Ma =+11. 96.
The phenylosazone
*
C.
Fischer and Morrell, Ber., 27, 382. Krauz, Ber., 43, 482; Z. Zuckerind.
Bohmen,
35, 570.
THE MONOSACCHARIDES
p-Rhodeohexose.*
633
CH
CHOH HCOH HCOH HOCH HOCH CHO

This sugar is formed from rhodeose in the same way as /3-rhamnohexose from rhamnose. The sugar has not been isolated in the pure crystalline form.
HEPTOSES
14
ALDOHEPTOSES
a-Glucoheptose.
CH OH HOCH HOCH HCOH HOCH HOCH CHO

2
acid.
The sugar is formed by reducing the lactone of a-glucoheptonic The sugar has been obtained by Fischer f in the form of large
to 190 C., and showing levorotation, melting at 180 19.7 (after solution = The sugar is slightly sweet and 25). is but slightly soluble in cold water (easily soluble in hot water) a-Glucocrystals
[a]
= D
heptose reduces Fehling's solution but to a less extent than d-glucose. Reduction with sodium amalgam gives inactive a-glucoheptite and oxidation with nitric acid gives inactive a-glucopentoxypimelic acid.
a-Glucoheptose-phenylosazone forms fine yellow needles melting at 195 C. * Krauz, Ber., 43, 482; Z. Zuckerind. Bohmen, 35, 570.
t Ann., 270, 64.
634
B-Glucoheptose.*
SUGAR ANALYSIS
CH OH HOCH HOCH HCOH HOCH HCOH CHO
2
acid,
This sugar is formed by reducing the lactone of /3-glucoheptonic but has not been isolated as yet in the crystalline form. Oxidation of /3-glucoheptose with nitric acid gives the dibasic 0-penis
toxypimelic acid whose lactone, CyHioOs,
dextrorotatory
([a] D
= +68.5).
/3-Glucoheptose-phenylhydrazone forms fine colorless needles melting at 190 to 193 C.; the phenylosazone of 0-glucoheptose is in every respect identical with that of a-glucoheptose.
d-Mannoheptose.
CH OH HOCH
2
HOCH HCOH HCOH CHOH CHO

d-Mannoheptose was obtained by Fischer and Passmoref from d-mannose by addition of hydrocyanic acid, and reduction of the lactone of the resulting d-mannoheptonic acid. The sugar was obtained in the form of fine needles, melting at 134 to 135 C.; it has a sweet taste, is The sugar is easily soluble in water but difficultly soluble in alcohol. dextrorotatory showing mutarotation, [a]g = + 85, 10 minutes after No perceptible fermentation was solution; [a]^ constant = + 68.64.
noted in presence of yeast.
Reduction of d-mannoIdentity of d-Mannoheptite and Perseite. heptose with sodium amalgam produces d-mannoheptite which was found by Fischer and Passmore to be identical with the natural heptite
alcohol perseite,
*
first
found by Avequinf in the

et Toxic., 7,
fruit of the alligator
Fischer, Ann., 270, 87.

t
t Ber., 23, 2226.
Ann. chem. med. Ph.
467 (1831).
THE MONOSACCHARIDES
pear (Per sea gratissima)
,
635
1888.
is
and
identified
relationship in properties of d-mannoheptite the following table (Fischer and Passmore)

:
by Maquenne* in and perseite
The
in
shown
636
SUGAR ANALYSIS
fermentable optically inactive sirup. Reduction of the sugar gave 1-mannoheptite melting at 203 C., which is higher than that observed for either of its components (187 C.). The same racemic compound was obtained by mixing equal parts of d-, and 1-mannoheptite
d,
and allowing the aqueous solution to

a-Galaheptose.
crystallize.
CH OH HOCH
2
HCJOH
H COH
HOC]
ino
a-Galaheptose was obtained by Fischer* from d-galactose by addition of hydrocyanic acid and reduction of the lactone of the resulting a-galaheptonic acid. The sugar was obtained only as a sweet, unferrnentable, levorotatory sirup.
a-Galaheptose-phenylhydrazone was obtained as fine colorless needles melting at 200 C. the phenylosazone consists of fine yellow needles, (m. p. 218 C.)
;
p-Galaheptose was obtained by Fischer* by reducing the lactone

of 0-galaheptonic acid, the latter being formed from d-galactose by addition of hydrocyanic acid at the same time as its isomer a-gala-
heptonic acid. excepting the

opposite in
The sugar has the same
structure as a-galaheptose,
and OH groups in the second carbon atom which are the two sugars; the particular arrangement belonging to
each sugar has not as yet been established. /3-Galaheptose crystallizes in the form of large prisms melting at 190 to 194 C. It has a sweet taste and is easily soluble in hot water.
levorotatory showing mutarotation, 54.4. minutes after solution) and [a]^ constant =
is
.
The sugar
[a] D
22.5 (10
This heptose sugar was obtained by Fischer f Volemose, C 7 Hi 4 7 by oxidation of the naturally occurring heptite alcohol volemite, which was discovered by BourquelotJ in the fungus Lactarius volemus.
*
Ann., 288, 139.
t Ber., 28, 1973. j Bull. Soc.
Mycol. de France,
5,
132;
Chem.
Ztg., 16, 190.
THE MONOSACCHARIDES
637
Volemose was obtained only in form of an impure sirup. The phenylosazone has the formula CrHtfCM^HCeHs^ and consists of yellow crystals melting at 196 C. The alcohol volemite CyRieO? consists of fine needles melting at 149 to 151 C., and showing in 10 per cent aqueous solution a dextrorotation of
[a] D
+ 1.92.
and volemite have not as yet been
configurations of volemose established.
The
KETOHEPTOSES
7 7 Hi 4 This ketoheptose was obtained by Bertrand* through the action The sugar was obtained in a of the sorbose bacterium upon perseite. 45 per cent of the perseite about the yield being pure crystalline form,
.
Perseulose.
taken.
Perseulose
[a]
is
after solution
strongly levorotatory and exhibits = 90 and [a]g constant = 81.
mutarotation,
of
Perseulose-phenylosazone, needles melting at 233 C.
CyH^CW^HCeH^,
consists
yellow
METHYLHEPTOSES
Rhamnoheptose.
CH
CHOH HCOH HOCH HOCH HCOH CHOH CHO
V.
This sugar was prepared by Fischer and Pilotyf by addition of hydrocyanic acid to a-rhamnohexose and reduction of the lactone of the resulting rhamnoheptonic acid. Rhamnoheptose was obtained only
as a colorless sweet sirup of
weak
dextrorotation,
is
([a] D
=+
8.4 about).
The phenylhydrazone
*
of
rhamnoheptose
rend., 147, 201;
characterized
by low
Compt.
149, 225.
t Ber., 23,
3102.
638
solubility in
SUGAR ANALYSIS
water and separates with great readiness. It consists of having the composition CgHieOe^HCeHs and melting
colorless needles
The phenylosazone, CsHuCMNsHCeHB^, consists of fine yelat 200 C. low needles difficultly soluble in water and hot alcohol; its melting point is about 200 C.
OCTOSES
C8 H
a-Glucooctose.
16
CH OH HOCH
2
HOCH HCOH HOCH HOCH CHOH CHO

This sugar was synthetized by Fischer* from a-glucoheptose the by addition of hydrocyanic acid yields 2 stereo-isomers, a- and
;
latter
/3-glucooctonic acid.
The lactone of a-glucooctonic acid gives upon reduction a-glucooctose. a-Glucooctose crystallizes from water in fine white needles as a
hydrate having the formula
Hi 6 O 8
[a]^
+ 2 H 0.
2
The sugar
is
levo-
rotatory and shows mutarotation;
50.5 (constant for the anall
hydride C 8 16 O 8 ). a-Glucooctose has a sweet taste and gives tion of a reducing sugar.
the ordinary reac-
a-Glucooctose-phenylhydrazone, CsHieOy^HCeHs, separates very readily as a difficultly soluble compound, consisting when pure of fine colorless needles melting at about 190 C. The phenylosazone forms
fine
position
to 212 C. and having the comHC6H5)2. Upon reduction with sodium amalgam a-glucooctose gives its alcohol a-glucooctite, C 8 Hi 8 8 which melts at 141 C.
yellow crystals melting at 210
C Hi 06(N
8 4
/3-Glucooctose, formed acid, has not been studied.

*
by reducing the lactone

Ann., 270, 64.
of |8-glucooctonic
THE MONOSACCHARIDES
d-Mannooctose.
639
CH OH HOCH HOCH HCOH HiOH HOH

2
CHOH CHO
This sugar was built up from d-mannoheptose by Fischer and Passmore * by addition of hydrocyanic acid and reducing the lactone of the The sugar was obtained only as a sweet resulting d-mannooctonic acid.
colorless
unfermentable sirup with slight levorotation, (W Z) = 3.3). d-Mannooctose upon reduction gives d-mannooctite CaHigOs, which
C.
consists of colorless very difficultly soluble crystals melting at 258
The phenylhydrazone CsHieOy^HCeHs forms colorless needles very insoluble in water and melting, when quickly heated, at about 212 C. The phenylosazone forms fine yellow needles very insoluble
in hot
water and alcohol, and melting at about 223 C.
a-Galaoctose.
CH OH
2
HOCH HCOH HCOH HOCH HOH HOH HO

a-Galaoctose was built up by Fischer f from a-galaheptose by adding hydrocyanic acid and reducing the lactone of the resulting
a-galaoctonic acid.
The sugar was obtained

O8
monohydrate C 8 H
levorotatory,
[a] D
]6
+ H O melting
2
at 109
as colorless crystals of the to 111 C. The sugar is
40 about.
*
Ber., 23, 2226.
t Ber., 27,
3198.
640
SUGAR ANALYSIS
sists of colorless
a-Galaoctose gives upon reduction a-galaoctite CsHisOs which conneedles melting at 220 to 225 C. The phenylhy-
drazone of a-galaoctose has the formula CsHieOT^HCeHs and melts to 205 C. The osazone GgHuOe^HCsHfi), forms fine yellow needles melting at 220 to 225 C.
at 200
METHYLOCTOSES
CH C H
3 8
15
Rhamnooctose.
CH CHOH HCOH HOCH HOCH HCOH CHOH CHOH CHO

3
This sugar was prepared by Fischer and Piloty* from rhamnoheptose by adding hydrocyanic acid and reducing the lactone of the The sugar was not separated in the pure resulting rhamnooctonic acid.
condition and
its
properties have not been determined.
NONOSES
CH
9
18
o-Glucononose.
CH OH HOCH HOCH HCOH HOCH HOCH

2
CHOH CHOH CHO

*
Ber., 23, 3102.
THE MONOSACCHARIDES
641
a-Glucononose was prepared by Fischer* from a-glucooctose by adding hydrocyanic acid and reducing the lactone of the resulting The sugar was obtained only as a colorless a-gluconononic acid. non-fermentable sirup with slight dextrorotation. Reduction of a-glucononose gives the alcohol a-glucononite, CgHÔg,
colorless crystals melting at 194 C. a-Glucononose-phenylhydrazone CgHigOs^HCeHs forms white needles only slightly soluble in cold water and alcohol and melting at about 194 C. The phenylosazone CgHuA^HCeHs^ consists of fine yellow needles almost insoluble in hot water and alcohol, and melting at 220 to 223 C.
which consists of
d-Mannononose.
CH OH HOCH HOCH Hci OH

2
HC<
HOH
d-Mannononose was prepared by Fischer and Passmoref from d-mannooctose by adding hydrocyanic acid and reducing the lactone of the resulting d-mannonononic acid. The sugar was obtained as white crystals melting at 130 C. and showing in aqueous solution dextrorotation ([]*[
=+50 about).
is
d -Mannononose
fermented by yeast with the same ease and com-
pleteness as d-glucose.
d-Mannononose-phenylhydrazone, CgHigOs^HCeHs, forms crystals The phenylosazone easily soluble in hot water and melting at 223 C. in hot water insoluble almost needles forms CgHisOr^HCeHj,^, yellow and alcohol and melting at 217 C. A peculiarity of d-mannononose is its striking resemblance to d-glucose.
The resemblance
and fermentability could
in composition, melting point, specific rotation easily cause confusion; an analysis of the osafix
zone easily serves, however, to

*
the class of the sugar (see page 371).
Ann., 270, 64. t Ber., 23, 2226.
642
SUGAR ANALYSIS
DECOSES
a-Glucodecose.
CH OH
2
HOCH HOCH HCOH HOCH HOCH CHOH CHOH CHOH CHO

This sugar has recently been prepared by Phillippe* from a-glucononose, following the usual method of adding hydrocyanic acid, saponifying and reducing the lactone of the resulting a-glucodeconic acid by means of sodiiim amalgam.
in
a-Glucodecose crystallizes in needle-shaped aqueous solution a dextrorotation, [a] '=
crystals,
which show
;
+ 50.4
(constant)
in
fresh solution [a]^ = 37. Under certain conditions the sugar may crystallize in plates having one molecule of water of crystallization. The sugar reduces Fehling's solution about 76 per cent as strongly as
glucose;
forms a phenylhydrazone melting at about 278 C. a-Glucodecose is reduced by sodium amalgam to the corresponding alcohol a-glucodecite, which consists of prismatic needles, melting and subliming at 222 C. and showing in aqueous solution [df
it
1.2.
*
Compt.
rend., 151, 986;
152, 1774.
CHAPTER XX
THE DISACCHARIDES
DIPENTOSE SACCHARIDES / C 5H 4 O x C H<A
9 5
disaccharide was obtained by Diarabinose, O'Sullivan* in heating Gedda gum with sulphuric acid (0.3 0.5 per Its is formation due to the down of cent). probably breaking higher condensation substances of the gum (arabinic acids) into diarabinose and other hydrolytic products. Diarabinose (also called arabinon,
CioHi 8
9.
This
arabiose and arabinobiose) was obtained by O'Sullivan as an amorphous vitreous hygroscopic substance of sweet taste and very soluble
in water
from which
198.8.
uct melts at about 75

[a] D
=+
precipitated by strong alcohol. The prodto 80 C., and is strongly dextrorotatory, It reduces Fehling's solution about 58.8 per cent as
it is
Analysis of the sugar and determination of its molecular weight by the freezing point method indicate the formula
strong as d-glucose.
Upon
heating with 2 per cent sulphuric acid, diarabinose
is
hydro-
lyzed quantitatively into 1-arabinose.

Diarabinose
1-Arabinoae.
PENTOSE-HEXOSE SACCHARIDES. /CH \

5 9
Glucoapiose,
CnH
2 oOio.
This disaccharide has not been isolated
as yet although its presence has been recognized by Vongerichtenf among the constituents of the glucoside apiin, which is obtained from The formation of glucoapiose from apiin should proceed acparsley.
cording to the following equation:

C* TJ f\ O26-tl 2 8<Ji4 Apiin
T
I
TT il 2 /"I
r^TTO VU-tlvlO
Glucoapiose
-4-
T^
f\eTTinOe îsp-ioysApigenm.
J.
Chem.
Soc., 57, 59;
59, 1029.
t Ber., 9, 1124;
33, 2334, 2904.
643
644
SUGAR ANALYSIS
glucoapiose, however,
is itself
The
into
decomposed by the hydrolytic agent

r^Ti/^ v^6-n 2 V-'6
-l
glucose and apiose
(p. 544).
CTT/^ Ill 0wio

2
_i_
~r
TT v/ Jl 2
_i_r^Tj^ T v^5-T-10^5>
Apiose.
Glucoapiose
d-Glucose
so that the separation of the disaccharide itself has not been accom-
plished
by
this
means.
.
This sugar has not been found as Galactoarabinose, CnH20 Oi It has been in nature. prepared synthetically by Ruff and Ollenyet
dorf* from ordinary lactose, by first oxidizing the sugar by means of bromine to lactobionic acid and then oxidizing the calcium salt of the latter with hydrogen peroxide in presence of basic ferric acetate; the COOH group of the acid is thus destroyed and a disaccharide sugar
obtained with 11
(
C atoms. CH OH
2
d-Galactose!
/nuYYtn
THE DISACCHARIDES
.
645
This glucoside has Methyl mannorhamnoside,* CiÂo CH 3 been obtained by hydrolysis of strophanthin, the poisonous principle of the seeds of Strophanthus Korribe, used by the natives of eastern Africa as an arrow poison. Strophanthin is decomposed by dilute
acids as follows:
=
Strophanthin
(C 27 H 380 7
H 0)
2
-f-
12
21
10
CH
3.
Strophanthidin
Methyl mannorhamnoside.
One part of strophanthin is dissolved in 5 parts of cold 0.5 per cent hydrochloric acid and then warmed for some time at 70 to 75 C. and then at 75 to 80 C. The strophanthidin which crystallizes out is
filtered off
and the cold
filtrate freed
from hydrochloric acid by means

is
of silver oxide.
The
clear filtered solution
then concentrated in a
vacuum
alcohol
is
to a sirup from which the methyl mannorhamnoside can be preThe compound upon recrystallizing from cipitated by means of ether.
is
obtained as white crystals melting at 207 C.
The
glucoside
easily soluble in water, fairly soluble in hot alcohol, but almost insoluble in ether. It is dextrorotatory ([a] D 8.24 about), unfermentable
=+
and does not reduce Fehling's solution. Upon heating with an excess of strong mineral acid, methyl mannorhamnoside yields large amounts of methylfurfural and levulinic acid. The glucoside is hydrolyzed by
heating with 5 parts of
1
per cent sulphuric acid as follows:

2
12
21
Oio
CH + 2 H
3
= C
12
C6 H O
12
+ CH OH.
3
Methyl mannorhamnoside
Mannose
Rhamnose
Methyl
alcohol.
DIHEXOSE SACCHARIDES
^ CeHiiO.
This group, by far the most important of the higher saccharides, includes the three well-known sugars sucrose, maltose and lactose.
:
SUCROSE.
Occurrence.
Saccharose.
Cane
Ci2
sugar.
22
On.
Sucrose occurs very widely distributed throughout the vegetable kingdom; from its importance as a commodity and food product it is the best known of the sugars. The approximate distribution of sucrose in different fresh plant
materials
is
as follows:
Percent.
Juice of green leaves ..................................... Juice of stalks from maize, sugar cane, etc .................. Sap of maple, birch, palm and other trees ............ ..... Apples, berries, oranges, prunes, bananas and other fruits ---.
0.1 2.0 1.0
2.0
20.0
5.0 14.0 12.0 15.0 25.0
Seeds, grains, nuts, etc ................................... Buds, blossoms and flowering organs ...................... ............... Roots, yams, bulbs, tubers, rhizomes, etc.
.
.
0.5 0.5 0.1 0.5
Feist, Ber., 31, 535;
33, 2063, 2069, 2091.
646
SUGAR ANALYSIS
Sucrose has not been identified with certainty in any products of purely animal origin. It occurs in honey in amounts ranging usually from 0.0 to 10 per cent; in abnormal cases the percentage of sucrose
exceed 10 per cent. The sucrose of honey, however, is derived primarily from floral nectar or other plant sources and must therefore
may
be regarded as of vegetable origin. Technical Processes. --The sugarcane, Preparation of Sucrose. sugar beet, maple, palm, sorghum and maize have all been utilized for the production of sugar. The annual production of raw sucrose for
about 16,000,000 long tons (1 long ton = 2240 Ibs.) of which about 8,500,000 tons are made from sugar cane and about 7,500,000 tons from the sugar beet; the production from other sources is insignificant. In the manufacture of raw sugar the juice is extracted from the sugar cane by means of mills, and from the sugar beet by means of diffusion batteries. The extracted juice is then clarified* usually with milk of lime, any excess of the latter being removed by means of carbon dioxide (" carbonatation "), sulphurous acid
the world at present
is
clarified juice,
is
The (" sulphitation "), phosphoric acid or other precipitating agent. which may contain from 10 to 18 per cent of sucrose,
In primitive countries the then evaporated to crystallization. is done in open pans directly over the fire; in the more modern factories some form of vacuum evaporator is used. After the evaporated juice has crystallized, the thick magma of crystals
evaporation
(" massecuite
"
or
"
fillmass ")
is
purged from
its
mother
liquor, or
molasses, a process which is usually carried out in centrifugals; the product thus obtained constitutes the raw sugar of commerce and
varies in purity
from 80 per cent to almost 100 per cent pure sucrose.
Refining.
The raw sugar
of
commerce
is
afterwards refined.
The
raw
process of refining comprises usually (1) washing the crystals of sugar with concentrated sirups to remove adhering molasses, a
process sometimes termed "affining," (2) dissolving the purified crystals in hot water and clarifying the solution with lime or other agents; (3) filtering the clarified solution over bone black f to remove coloring
matter and other impurities; (4) evaporating the filtered and decolorized solution to a magma of crystals; (5) centrifuging the "masse" " cuite or fillmass" and drying the pure white crystals of sucrose in
of substances which have been proposed for clarifying sugar almost unlimited. A classification of clarifying agents made by Lippmann (Die Deutsche Zuckerind., 34, 9) includes 620 different materials or processes. t The use of bone black has been largely discontinued in the refining of beet
is
The number
juices
sugar.
THE DISACCHARIDES
the demands of the trade.
647
granulators, or in cones, cubes, dominos or other forms according to Refined sugar is usually about 99.8 to
99.9 per cent pure, the remaining 0.1 to 0.2 per cent consisting mostly of moisture with occasional traces of ash, invert sugar, raffinose and cara-
mel substances.
obtain sucrose perfectly pure the best grade of refined sugar is The method recrystallized from neutral redistilled 96 per cent alcohol. described upon page 121 may be used to advantage.
To
For the separation of Isolation of Sucrose from Plant Substances. sucrose from plant substances, when only small amounts of the sugar are present, Schulze* has made use of the difficultly soluble strontium
The fresh material, in presence of an excarbonate to neutralize any acidity, calcium powdered After cooling, the extract is is extracted with hot 90 per cent alcohol. filtered and then heated to boiling with the addition of a hot saturated
bisaccharate
C^HÔn
2 SrO.
cess of pure finely
solution of strontium hydrate using over 3 parts of Sr(OH) 2 for every After boiling 30 minutes the 1 part of sucrose supposed to be present. and again boiled for 30 alcohol with is washed filtered, precipitate
minutes with strontium hydrate solution. The precipitate is filtered hot, using a hot water funnel, and then, after suspending in water, decomposed with a stream of carbon dioxide. The solution is filtered from strontium carbonate and then evaporated to a sirup which is purified by
means of neutral 95 per cent alcohol in the usual way. The alcoholic solution is reevaporated to a sirup and repurified as before, the process of evaporation and extraction of the sirup with alcohol being repeated several times. The final sirup is placed over concentrated sulphuric acid
in a cool place for crystallization.
PROPERTIES OF SUCROSE
Crystalline Form.
Sucrose crystallizes in beautiful colorless crys-
Fig. 195.
Monoclinic crystals of sucrose. I, Tabular form; hemihedral faces.
II,
Form with
tals
surfaces
have hemihedral belonging to the monoclinic system. The crystals and show the greatest variety of form (Fig. 195). The shape
* Z. Ver.
648
SUGAR ANALYSIS
of sucrose crystals is greatly modified by other substances, the effect of raffinose in this respect being especially pronounced (p. 735). Crystals of sucrose may take up soluble coloring matter from the mother
liquor during growth and such crystals often show a variation in tint when viewed in different directions (pleochroism) Although sucrose in solution is optically active, its crystals, as was first noted by Biot,* do
.
not rotate the plane of polarized light. Melting Point and Specific Gravity.
is
The melting point
of sucrose
between 160 to 180 C., the given by variations being due apparently to differences in method and in the
different observers as varying
The specific gravity of solid sucrose physical character of the sugar. is given by. different authorities as between 1.58 and 1.61, the differences being probably due to variations in the character of the crystals. The recent determinations of Plato f give for chemically pure sucrose
j|s
=1.591.
The
is
specific gravity of the hypothetical solid sucrose in
= 1.55626; the difference given by Plato as between this figure and that for the actual solid being due to the conaqueous solution
traction in
d^
'
volume during
(p. 33).
solution.
plays in the determination of sucrose
The part which this phenomenon by densimetric methods has already
been considered
Solubility.
The
peratures and the character
solubility of sucrose in water of different temof the solutions thus obtained are given by
HerzfeldJ in Table XCI. Sucrose is soluble in 80 parts of boiling absolute alcohol, more
easily soluble in dilute alcohol
but insoluble in ether.
SOLUBILITY OF SUCROSE AND THE MELASSIGENIC ACTION OF SALTS
The solubility of sucrose, as of all other sugars, is affected to a marked degree by the presence of foreign organic and inorganic substances.
Such
impurities play an important part technically in preventsatuing the recovery of sucrose from sugar-house molasses. rated solution of sucrose in contact with sucrose crystals can dissolve
no more sucrose at constant temperature; if solid potassium acetate, or sodium chloride, or many other salts be stirred into the solution, however, it will not only be dissolved but more of the sucrose will also enter solution. In other words more sugar will be dissolved than can be held in solution by the water alone. This phenomenon is explained by many authorities as being due to the formation of sucrose-salt com*
M6moires de l'Acad6mie,
13, 59, 126.
| Z. Ver.
f Z. Ver. Deut. Zuckerind., 50, 1012. Deut. Zuckerind., 42, 181, 232.
THE DISACCHARIDES
alone.
649
pounds, or complexes, which have a greater solubility than the sucrose
TABLE XCI.
Solubility of Sucrose in Water at Different Temperatures.
Temperature.
650
SUGAR ANALYSIS
This process has given place technisolution of undialyzed impurities. sucrose of to the saccharate recovery to be described later. process cally In low-grade sugarSolubility of Sucrose in Cane Molasses. cane molasses an opposite condition exists to that in beet molasses; in cane molasses the amount of sucrose is less than that which will saturate the quantity of water present.
This
is
shown by the following

Per cent.
analysis of a low-grade cane molasses.
Water
Sucrose Invert sugar
Salts
20 30 30
8 12
Organic non-sugars
Geerligs's
Theory
of Melassigenic Action.;
molasses of the
above composition can dissolve no more sucrose, yet the 20 parts of water alone could hold in solution 40 parts of sucrose at ordinary temThis difference in behavior upon the part of cane molasses perature. * is explained by Prinsen Geerligs as due to a combination between the molasses invert sugar and the salts of the (the potassium organic salts more especially). The invert-sugar-salt complexes which are thus in formed hold combination a large amount of water of hydration which thus reduces the quantity of water available for solution of the The power of sucrose to form salt complexes is much less sucrose. than that of invert sugar so that it is only in cane molasses of very low invert sugar content that sucrose-salt complexes exist in sufficient quantity to raise the solubility of sucrose above the saturation point of the water present.
According to this theory the addition of anhydrous glucose to a saturated solution of a sucrose-salt complex should displace the sucrose
and cause a part of the latter to crystallize out. This was verified experimentally by Geerligs who found that when 225 gms. of anhydrous glucose were added to 300 gms. of a saturated solution of sucrose and potassium acetate, and the mixture allowed to stand for several months
75 gms. of sucrose separated in the crystalline form. A check solution without addition of glucose showed no evidence of crystallization. The melassigenic action of different organic and mineral substances
plete review of the various physical subject the student is referred to the
* t t
upon sucrose has been studied by many investigators and for a comand chemical theories upon the works of Lippmann f and GeerZ. Ver. Deut. Zuckerind, 45, 320.
"
"
Chemie der Zuckerarten," 1147-1162. Cane Sugar and its Manufacture " (1909), 301-317.
THE DISACCHARIDES
651
The boiling point of aqueous Boiling Point of Sucrose Solutions. sucrose solutions of different concentrations is given by Gerlach * as
follows
:
Per cent sucrose

Boiling point
....
10 20 30 40 50 60 100.4 100.6 101.0 101.5 102.0 103.0
70
80
90 8
106.5 112.0 130.0
Specific Rotation.
The
specific rotation of sucrose
has been the
subject of greater study than that of any other sugar. The first determinations were made in 1819 by Biot,f who first introduced the constant of specific rotation and thereby founded the science of optical
analysis.
The value
very closely
for the specific rotation of sucrose in

2
[a]
=+
aqueous solution
is
66.5.
tion, solvents, salts, etc.,
The influences of temperature, concentraupon the specific rotation of sucrose have
Alcoholic Fermentation. In so far as the various yeasts, moulds and bacteria secrete the enzyme invertase they are able to ferment sucrose in the same manner as its products of The majority of the yeasts secrete ininversion, glucose and fructose.
vertase and ferment sucrose with the
already been considered. Fermentations of Sucrose.
same
yield of alcohol
and carbon
is
dioxide as
is
obtained from glucose and fructose; the process
some-
what
slower, however, in its first stages owing to the retarding effect of the inversion which must precede fermentation.
A considerable number of Non-inverting Yeasts and Moulds. alcohol-producing organisms, such as Saccharomyces octosporus,l Saccharomyces apiculatus, and most of the Mucor genus of moulds do
not secrete invertase and pure cultures of these do not ferment sucrose. Attempts have been made to employ organisms of this class such, for example, as Mucor circinelloides,\\ for destroying the invert sugar of cane molasses, in the hope of obtaining the residual sucrose in a suitable condition for recovery.
success. |f
* f
The
process has not been a technical
Z. Ver.
Deut. Zuckerind,
13, 283.
II
Memoires de 1' Academic, 2, 41; 13, 118. Fischer and Lindner, Ber., 28, 984. Fischer and Lindner, Ber., 28, 3034. Gayon, Ann. chim. phys. [3], 14, 258.
f Upon the basis of Prinsen Geerligs's molasses theory it is evident that fermenting away the invert sugar of cane molasses would have but little effect upon rendering the sucrose more crystallizable. The result would simply be to change
the molasses from a cane to a beet type. Suppose a cane molasses of the following composition to have its invert sugar fermented and the solution of sucrose, salts
652
Lactic
SUGAR ANALYSIS
and Butyric Fermentations.
The
lactic
and butyric acid
fer-
mentations can be produced with sucrose by the same organisms which produce these fermentations with d-glucose and d-fructose. In a few cases, however, where the particular organism does not secrete invertase, fermentation of sucrose does not take place.
Fig. 196.
terioides.
Leuconostoc mesen-
Fig. 197.
After Zopf. (48hour culture in molasses showing slimy envelopes of dextran .)
Bacterium pediculatum. Koch and Hosaeus.
After
Viscous Fermentation.
One
of the
most common fermentations of
sucrose observed in sugar factory experience is the so-called viscous fermentation by which sucrose is converted into the gum dextran.
The
have:
best
known dextran-producing organism

to
is
the Leuconostoc mesen-
and non-sugars
be evaporated to the original concentration,
We
would then
THE DISACCHARIDES
terioides (Fig. 196),
653
earliest investigators
which was supposed by the
to ferment sucrose according to the following equation:
C H On = C
12
22
10
+ CH
6
12
6.
Sucrose
Dextran
Fructose.
Later researches * showed, however, that the action of Leuconostoc and of many other " dextran-formers " consisted first in an inversion of the sucrose into d-glucose and d-fructose so that the above formula is
not strictly correct;
it was also established that dextran is a polysaccharide (C 6 Hio0 5 )n and that it constitutes the slimy jelly-like capsule in which the organisms are embedded. The dextran is, therefore, to
be regarded of assimilative, rather than of fermentative (i.e., enzymic) origin. Very similar to Leuconostoc in its action is the Bacterium pediculatum discovered by A. Koch and Hosaeus in the sirup of a
sugar factory.
The organism secretes a slimy capsule of gum which, becoming greatly elongated upon one side, gives it a stem-like appearance (Fig. 197). Certain gum-producing organisms have been found, such as Micrococus gelatigenosus, Bacillus gwnmosus, Bacterium gummosum, etc.,
which form dextran from sucrose but not from glucose. This has been regarded as a fermentation of sucrose without preceding inversion; most of the members of this class of organisms are found, however, to secrete invertase so that the sucrose in these, and no doubt in all other cases, where fermentation or assimilation takes place, is probably first inverted. The peculiarity which certain bacteria have of forming dextran from sucrose but not from glucose may be explained by supposing that these organisms are able to ferment or assimilate glucose only at the time of its separation from the sucrose molecule (i.e., in its nascent Even in the case of Leuconstate) and not after it is already formed. ostoc, which can assimilate free glucose and fructose, the formation of
is several times more rapid with sucrose. f In the so-called visFormation of Mannite During Fermentation. cous or gummy fermentation of sucrose mannite is frequently formed
dextran
MonoyerJ regarded the two substances as products of separate fermentations which he formulated as follows:
in addition to dextran.
Mannitic fermentation:
13 Ci 2
*
22
On
+ 25 H
24
C H 14
6
6 -f-
12
C0
2.
(1)
Sucrose
Mannite.
of
For a
full
account of the action of Leuconostoc upon sucrose see the work
Liesenberg and Zopf, Centralbl. fur Bakteriologie, 12, 659; 13, 339. " " Cane Sugar and its Manufacture (1909), 38. t Prinsen Geerligs 1862. | These pour le doctorat en medecine, Strasbourg,
654
SUGAR ANALYSIS
fermentation:
12 Cis
Gum
12
Gum.
+ 12 H 0.
2
(2)
According to the above combined equations 100 parts of sucrose gum and 51.1 parts of mannite. This proportion is not fixed, however, the variation in yield being explained by the predominence of one or the other fermentation. It is more probable, however, that the dextran is formed as an assimilative and the mannite
yield 45.5 parts of
as a reduction product in
species of bacteria.
many
anaerobic fermentations by a single
The gum, which

crose,
(p.
is
is
produced in the viscous fermentation of suIt
not always dextran.
may
also consist of levulan or levan

is
615),
which give fructose upon hydrolysis, whereas dextran
hydrolyzed only into glucose. Influence of Fermentation

of Sucrose.
Gums Upon
Polarimetric Determination
gums
and levorotatory products may introduce a considerable error in the Browne* reported the following polarimetric estimation of sucrose. analyses of badly fermented sugar-cane juices:
of highly dextrorotatory
in sugar-house
Degrees Brix.
The presence
THE DISACCHARIDES
lose,
655
weighing in some cases several pounds. The product after purifying with hot sodium hydroxide was colored blue with zinc chloride and iodine, was soluble in ammoniacal copper solution and had the composi-
The amount of cellulose formed by the organism was about 7 per cent of the total sucrose destroyed.
tion of cellulose.
The citric fermentation (p. 585) may also Citric Fermentation. occur with sucrose, the fungus Citromyces glaber yielding 50 per cent A Citromyces found by Browne * upon hotof the sucrose in citric acid.
room molasses in Louisiana was found to contain over 11 per cent chitin; the latter gave upon hydrolysis with hydrochloric acid over 60 per cent
of pure glucosamine chloride (p. 753).
mentioned,
lactic
Among other fermentation products of sucrose, besides those already may be mentioned butyl and amyl alcohols and acetaldevaleric,
hyde; formic, acetic, butyric, propionic,
capronic,
caprylic,
and succinic acids; as well as the gaseous products hydrogen and methane. For a description of the fermentations which give rise to these and other substances the student is referred to the works of Lafar,f JorgensenJ and Lippmann.
DECOMPOSITION OF SUCROSE BY HEATING

Sucrose upon heating above its melting point begins to decompose with evolution of water. Between 170 and 190 C a mixture of brownish colored substances, known as caramel, is formed; above 190 C.
large quantities of carbon dioxide and monoxide are given off together with various volatile decomposition products such as aldehyde, acetone,
From acrolein, furfural and even benzolderivatives, as benzaldehyde. a technical viewpoint the most important of these decomposition
products
is
caramel.
Caramel is usually prepared by heating sucrose to 170 to 190 C. and consists of a mixture of decomposition products, the exact compoThe caramelization or sition of which has not been fully ascertained.
browning of sucrose
may
begin, however, at temperatures below 100
in presence of moisture.
As ordinarily prepared from sucrose caramel
consists of a brownish colored substance, easily soluble in water but in-
Caramel reduces soluble in strong ethyl alcohol, ether or chloroform. from solution it is solution precipitated completely strongly; Fehling's by ammoniacal lead subacetate. Solutions of caramel show before the
*
J.
Am. Chem.
"
Soc., 28, 465.
t Lafar's t
Technische Mykologie," Jena (1901-1907). " Mikroorganismen der Garungsindustrie," Berlin. Jorgensen's " Chemie der Zuckerarten," 1288-1317.
656
SUGAR ANALYSIS
spectroscope characteristic absorption bands, the blue part of the
spectrum being more or less extinguished according to concentration. If a caramel solution is shaken with an alcoholic solution of paraldehyde and allowed to stand in the cold for 24 hours a brownish yellow
gummy
upon pound
precipitate will form, the rapidity of deposition depending the amount of caramel present. The paraldehyde-caramel comis
from which it is reprecipitated by strong not been definitely established. has composition alcohol; The formation of caramel from sucrose consists primarily in the
soluble in water
its
splitting off of
water in successive stages, this giving
rise to
a series of
dehydration and condensation products of varying complexity. Gelis* was the first to attempt the separation of caramel into its components and defined three different constituents, caramelane, caramelene and carameline. Caramelane was prepared by Gelis by heating sucrose until it lost about 12 per cent in weight and was given the formula C^HigOg; caramelene, C^HÔu H 2 0, was prepared by heating sucrose until it lost about 15 per cent in weight; and carameline, CgeHiooCô H 2 0, by heating sucrose until it lost about 20 per cent in weight. Other investigators give caramelane, caramelene and carameline entirely different formulae;
each of these substances
is
composition products so that the formulae assigned questionable value.
probably a mixture of deby Gelis have a
Caramelane was prepared by Stollef by heating melted sucrose at 180 C. until no further loss occurred; the residue was dissolved in water, any unchanged sugar removed by fermentation and the residue
evaporated in vacuo to dry ness. The substance thus obtained consisted of a brownish mass melting at 134 to 136 C., its composition corresponded to the formula C^HisOg the same as the caramelane of
Gelis.
saccharan, Ci 2 Hi 8 9 obtained by Ehrlich by sucrose to It is 200 heating C., has already been described (p. 467). with the identical caramelane of Gelis and Stolle. probably
,
The caramel substance
Destructive Action of Heat
Upon Sucrose
in Solution.
A knowl-
edge of the destructive changes which sucrose undergoes upon heating its aqueous solutions is of great importance. Unfortunately no fixed rule can be given for this, as the nature and extent of the decomposition
depend largely upon the character of accompanying impurities.
Sucrose in perfectly neutral solutions, when heated for a few hours at 100 C., begins to undergo decomposition as a result of carameliza*
t Z. Ver.
Ann. chim. phys. [3] 52, 352; Compt. rend., 46, 590. Deut. Zuckerind, 49, 800; 51, 836; 53, 11-47.
THE DISACCHARIDES
tion
657
and incipient inversion, the latter being produced according to some chemists by the H ions of dissociated molecules of water, and according to other chemists by auto-inversion, the sucrose itself behaving After the commencement of inversion as an extremely weak acid. the sucrose solution becomes perceptibly acid, and heating from this point causes decomposition and inversion to proceed with increasing
rapidity.
To determine
upon heating
its
solutions
the rate of decomposition which sucrose undergoes when formation of free acid is prevented,
Herzfeld* conducted
experiments with solutions which were
made
slightly alkaline; variations in the kind and amount of alkali were not found to cause any difference in the character of the results. The fol-
lowing table taken from Herzfeld's work shows the percentage loss of total sucrose caused by heating solutions of different concentration at
varying temperatures for
hour.
TABLE XCII
Loss of Sucrose upon Heating Solutions of Different Concentration at Varying Temperatures
Deg. C.
658
For the
is
SUGAR ANALYSIS
first few hours of heating only a slight decrease in polarizanoted, then, with the formation of acid oxidation products and the consequent increase in the rate of inversion, the polarization quickly
tion
falls until
its
inversion
etc.),
the polarization of undecomposed sucrose, and that of and decomposition products (glucose, fructose, caramel, exactly neutralize one another and the rotation is 0. Upon
at
longer heating the remaining sucrose is inverted; the rotation of the fructose becomes the predominant factor and the polarization is levoA maximum levorotation is reached at C, after which, with rotatory.
48^56
64
72
80
88
96
Hours of Heating
Fig. 198.
Showing changes
in polarization of
a sucrose solution by destructive
action of heat.
the decomposition of the more unstable fructose, the rotation again until at a second point of inactivity is reached, the approaches rotatory powers of undecomposed fructose, glucose and other sub-
stances counterbalancing one another. Upon longer heating the remaining fructose is destroyed; the rotation of glucose is now the predominant factor and the polarization of the solution becomes dextrorotatory again. A maximum dextrorotation is reached at E, after which with the destruction of the glucose the polarization gradually
approaches 0, until at F a third and final point of inactivity is reached. The curve of changes just described may be longer or more contracted than that shown in Fig. 198 according to the temperature of heating, nature of salts and impurities present, and other conditions.
High-polarizing Sugar.
condition exactly opposite to that just
noted
is
sometimes observed in technical operations, where heating
THE DISACCHARIDES
to cause
659
concentrated sucrose solutions has been found under certain conditions an increase in the polarization. This phenomenon has been
attributed by some to the formation of high-rotating dextrinoid condensation products and by others to the splitting off of glucose in a high mutarotating form. This increase in polarization, according to Lippmann,* is observed only when the solution is neutral or very
free alkali it does not seem to take place. If sucrose is heated with only a small Optically Inactive Sugar. amount of water at 150 to 160 C. for a short time, a mixture is ob-
weakly acid; in presence of
tained which shows almost complete optical inactivity. This so-called optically inactive, or neutral sugar, was first observed by Berzelius
and Mitscherlich,f and has been the subject of frequent investigations since their time.
Optically inactive sugar consists of a mixture
of glucose
and other products whose rotations neutralize one another. According to Wohlf the sucrose is decomposed into glucose and a condensation product of fructose which he calls levulosin.
n (CwHaOu)
Sucrose
= nC6 H 12
Glucose
(C 6 H 10 O5 )n.
Levulosin.
[0^=4-66.5
~~H^=o.
Optically inactive sugar upon warming with acids becomes strongly levorotatory and this is explained by the hydrolysis of the levulosin
into d-fructose.
(C 6 Hio0 5 )n
Levulosin
+nH
= n C Hi
6
6.
d-Fructose.
THE INVERSION OF SUCROSE

INVERSION OF SUCROSE BY ACIDS
One of the earliest facts noted in connecEarly Investigations. tion with the chemistry of sucrose was that after warming with acids
the sugar could no longer be recovered in its original crystallizable form. The change was described by saying that the sucrose was converted into "uncrystallizable sugar," a term which is still occasionally used by certain writers. After the invention of the polariscope, Biot,
noted that the change which acids produced upon sucrose was attended by an alteration in the character of the rotation imparted to the plane of polarized light; the direction of rotation for the original
in 1836,
sucrose solution
to On was changed from right to left, or from " " account of this transposition in sign the term inversion was applied
.
* "
Chemie der Zuckerarten," 1223;

pharm.
[3],
Z. Ver. Deut. Zuckerind, 35, 434.
t Journ.
4, 216.
% Ber., 23, 2088.
660
i
SUGAR ANALYSIS
and the name " invert-sugar " given to the products It was soon observed that the sirupy sugar obtained
to the process of the reaction.
by inverting sucrose soon crystallized with separation of glucose; Dubrunfaut,* however, was the first to explain the true character of the process and showed that the sugars glucose and fructose were both formed during inversion. It was noted quite early in Wilhelmy's Law of Mass Action.
the study of inversion that the various acids differed in the rapidity of their inverting power, although the action of each acid seemed to fol-
low one general law. The nature of this law was discovered in 1850 by Wilhelmy,t who showed that the amount of sucrose inverted by an acid in a given moment of time is always a constant percentage of the
amount
This discovery is formulated in the velocity of a reaction at any moment is proportional to the concentration of the reacting substance. The inversion of sucrose by acids is expressed by the equation:
of
unchanged sucrose present.

viz.,
Wilhelmy's law of mass action;
12
Sucrose
H On + H = C H Water Glucose
22
2 6 12
+CH
6
12
6.
Fructose.
Although this equation involves the disappearance of one molecule of water with each molecule of sucrose, and is therefore bimolecular, the diminution in the total active mass of water is so slight that the process
of inversion can be treated as a unimolecularf reaction. Rate of Inversion. If a is the original amount of sucrose present and x the quantity inverted at the end of the time t after the com-
mencement
of inversion, then the rate of inversion for a unimolecular reaction will be:
S = *(a-*),
in
(1)
which dx
is
the infinitesimal quantity of sucrose inverted during the
infinitesimal period of time dt and k the velocity coefficient of the inversion. The constant k is found by means of the integral calculus to be
fc
lognat.
(2)
or,
changing from natural to

(log nat.
common
logarithms,
-T-
log com.
0.4343).
*
t
Compt.
rend., 42, 901; 69, 438.
Poggend. Ann., 81, 413, 499. " t For a demonstration of this see Mellor's Chemical Statics and Dynamics" (1909), pp. 40 and 41.
THE DISACCHARIDES
661
For purposes of comparison, however, formula (2) using common logarithms is often employed. Determination of Rate of Inversion by Polariscope. In applying formula (2) the polarimetric observations may be substituted for a and x. Calling the rotation before inversion r and after inversion r
M
and
for
any time
during inversion
r,
then
oo
following table shows the rate of inversion at 20 C. normal weight (26 gms.) of sucrose made up with water and 10
The
for"
c.c. of
concentrated hydrochloric acid to 100 true
c.c.
TABLE XCIII
Showing Rate of Inversion of Sucrose
662
SUGAR ANALYSIS
The somewhat
often obtained
sion,
Errors in Polarimetric Method for Determining Rate of Inversion. irregular values for the velocity coefficient, which are
by the polarimetric method at the beginning
of inverof
have led some investigators to suspect an exception to the law

for the early stages of hydrolysis.
mass action
The method
of deter-
mining the rate of inversion by observing the changes in the rotation in a polariscope tube is attended with several small There is, first, the possible influence of the contraction f in errors.* volume which accompanies inversion, and which for a 25 per cent There is, second, the solution of sucrose is about 0.5 c.c. per 100 c.c. change in polarization of the liberated glucose and fructose due to mutarotation, this error, however, being greatly reduced by the accelof a solution
The supposition that the increase in erating influence of the acid. concentration of fructose during inversion causes an error in the value
been proved by Rosanoff. Clark and Sibley to be untrue, since the percentage of water in the solution remains practically constant during the inversion. Careful experiments by the above authorities,
of k has
in
which varying amounts of sucrose were inverted in solutions con-
taining the same weights of acid and water per unit volume, show that the velocity coefficient is independent of the initial concentration of
the same throughout inversion as long as the concentraand acid remains unchanged. Another source of error in measuring the constant k is due to the slight rise in temperature which takes place in mixing the acid and sugar
sucrose and
is
tion of water
The speed of inversion is thus slightly accelerated at the beginning and this would explain the slightly higher values of k for the first few readings of the previous table.
solution.
Inverting
Power
of Different Acids.
The
inverting power of
different acids has
been determined by Ostwaldf whose results are
given in the following table. To avoid the use of small decimals the constant C = 10,000 k is employed. The second column of the table gives the relative inverting power of each acid as compared with
that of hydrochloric acid .which is taken as 100. In making the experiments 10 c.c. of a 40 to 50 per cent sucrose solution were inverted at
25 C. with 10
c.c.
of a
normal solution of the
acid.
* For a fuller discussion of these errors see paper by C. S. Hudson (J. Am. Chem. Soc., 32, 885), " Is the hydrolysis of cane sugar by acids a unimolecular reaction when observed with a polariscope? " and the paper by Rosanoff, Clark and Sibley (J. Am. Chem. Soc., 33, 1911), "A reinvestigation of the velocity of sugar
hydrolysis."
t
Lippmann's
"
Chemie der Zuckerarten
"
p. 1258.
J J. prakt.
Chem.
[2],
29, 385.
THE DISACCHARIDES
TABLE XCIV
Showing
Relative Inverting
663
Power
of Different
Adds
Kind
of acid.
664
solution.
SUGAR ANALYSIS
The formula
Ci 2
for the inversion of sucrose
is
in fact
sometimes
written:
22 Oii
Sucrose
HO+ + Water
2
H ion
H=
12
Invert sugar
+HH,
ion,
ion participating in an unlimited number of reactions. Many hypotheses have been proposed to account for the catalytic action per-
one
formed by the
yet been found.
action, carrier of water, etc.,
during the inversion of sucrose, such as vibratory but no satisfactory explanation has as
Elevation of Upon Speed of Inversion. increase in the a marked inverting power of temperature produces k coefficient about 15 the velocity increasing per cent for each acids,
Influence of Temperature
This rate of increase, which is approximately the same however, with rise in temperature; the k in from to 10 C. was found by Hammerschmidt * increase total to be about 500 per cent, from 30 to 40 C. about 400 per cent and
1
C. elevation.
for all acids, diminishes,
from 70 to 80 C. about 300 per cent. Arrhenius's Hypothesis of "Active" and "Inactive" Sucrose Molecules. Inasmuch as the ionization of acids in aqueous solution is not greatly affected by changes in temperature and as the coefficient for the increase
in speed of the
H ions for 1
amount
"
of
C. increase
is
only a small percentage of the
increase observed for the inversion constant k, Arrheniusf adopted the " " hypothesis that solutions of sucrose contained active" and inactive"
molecules, the
"
compared with the
inactive," but
active sucrose" being relatively small, as this amount increasing, at the
" expense of the inactive" sucrose, by about 12 per cent for each 1 C. This transformation of "inactive" into "active" sucrose preincrease.
cedes inversion and

or
is
supposed to take place through addition of water
of molecular rearrangement. Upon this hypothesis Arrhenius has derived the following formula for expressing the influence of temperature
by some process
upon the inversion
of sucrose
between
and 55 C.
in
which
Cti
and
Cto are the inversion coefficients of the acid at the
tem-
peratures ti and to, TI and T being the corresponding temperatures in absolute degrees; e is the constant 2.71828 (the natural logarithmic
base) and q is the thermal constant for the transformation of "inactive" into "active" sucrose which is estimated to be 25,600 calories
*
t Z. physik.
Chem.,
4, 227.
THE DISACCHARIDES
665
" inactive" sucrose. This formula of Arrhenius per gram molecule of according to Ley* also holds for temperatures above 55 C. Of other hypotheses, which have been Hypothesis of Sucrose Ions.
may
proposed to explain the effect of temperature upon inversion velocity, be mentioned the so-called "acid nature" of sucrose in accordance with which sucrose is supposed to become dissociated into ions. The formation of saccharates or salts of sucrose is used as one argu-
for this hypothesis; solutions of sucrose, however, show perfect neutrality to the most sensitive indicators, and are absolute non-con-
ment
ductors of electricity, so that no direct evidence exists to support the ions. hypothesis of sucrose
Influence of Concentration and of Salts Upon Inverting Power of The inversion velocity of sucrose by means of acids is in genAcids. eral proportionate to the concentration of ions; strict conformity to
this rule, however, obtains only with pure dilute solutions of the acid. The proportionality of the inversion constant k to concentration of
H ions
shows marked deviations at high concentrations of acid or in

salts.
presence of neutral
tration of 0.1
Thus the proportionality

nitric acid is
in
H ion
but
concen1: 4.64;
normal to 0.5 normal
not
1: 5
:
the proportionality in inverting power, however, is 1 6.07. This increase in the proportionality of the inversion constant is explained by In the same way addition of ions. an increase in the speed of the
ions, potassium nitrate to nitric acid will lower the concentration of but cause an increase in inversion velocity, this increase being explained by the increase in speed imparted by the dissociated molecules of potas-
sium nitrate to the remaining H The observations just noted

for
ions.
for nitric acid
and potassium
nitrate
the strong acids and their corresponding hold, however, only neutral salts. With weak acids an exactly opposite effect is noted. Increasing the concentration of acetic acid, for example, lowers the
proportionality of the inversion constant fc; so also the addition of an equivalent amount of potassium, acetate to acetic acid will reduce the
value of k to
$ of its original
amount.
Additions of neutral salts of a different acid than the inverting Thus sodium sulphate diminishes agent produce variable effects.
while sodium chloride increases the inversion velocity of acetic acid. In addition to the view that neutral salts alter the activity of the " " is active sucrose ions, Arrhenius supposes that the amount of also affected, while other chemists hold that the molecules of water
undergo dissociation to a greater or

* Z. physik.
less degree.
Chem., 30, 253.
666
SUGAR ANALYSIS
Organic non-conductors, such as alcohol, acetone, etc., if present in large amounts, diminish the inversion velocity of acids to a marked degree, although the electric conductivity of the solution itself may not be appreciably lessened. In such cases it is supposed that the
movement
of the
H ions
is
in
some way retarded.
Further discussion of the numerous hypotheses which have been proposed in this connection must be passed over; for a fuller treat-
by acids and the relationship of the dissociation the student is referred to Lippmann,* the to theory subject or to the more special treatises upon physical chemistry.
ment
of the inversion of sucrose
INVERSION OF SUCROSE BY SALTS

Sucrose is inverted upon heating with solutions of metallic salts; the speed of inversion, as in the case of acids, is in general proportionate to the concentration of hydrogen ions, the latter being formed by a hydrolysis of the salt in presence of water according to the following
equation
MA + HOH Water
Salt
MOH
Hydroxide
H-A,
Dissociated acid
The concentration of is the metal and A the acid radical. which and hence the speed of inversion, depends upon the extent of hydrolysis and dissociation. A number of investigators have studied the inversion of sucrose by salts. Walker and Aston,f working with sucrose solutions at 80 C., found the following inversion constants for a number of nitrates
in
H ions,
Cadmium nitrate (N/2) Zinc nitrate (AT/2) Lead nitrate (N/2) Aluminum nitrate (N/2)
0.000154 0.000207 0.001590 0.007700
The same
investigators.
order, Cd, Zn,
Pb and Al has also been found by other Long,t who has made an extensive study of the in;
verting action of salts, found for several sulphates the inversion to increase in the order Mn, Zn, Fe and Al. Kahlenberg, Davis and
from a study of the inverting power of different salts at C. (the temperature of boiling acetone) by the polariscopic and freezing-point methods obtained the following results:
Fowler
55.5
*
"
t J. t
Chemie der Zuckerarten," 1257-1303. Chem. Soc., 67, 576. J. Am. Chem. Soc., 18, 120, 693. J. Am. Chem. Soc., 21, 1.
THE DISACCHARIDES
667
Salt.
668
chloride,
SUGAR ANALYSIS
and
5, 10,
20 and 30 per cent invert sugar were heated to 100

5 7.47
for 3 hours.
Per cent invert sugar Per cent sucrose inverted
10 15.05
is
20
21.93
30
27.50
The
influence of different salts of the
same acid
shown
in the fol-
lowing series, where 50 c.c. of solutions containing 40 per cent sucrose and 25 per cent invert sugar were heated at 100 C. for 2 hours with a quantity of different chlorides equivalent to 1.75 gm. Cl. KC1 NaCl LiCl CaCl 2 SrCl 2 BaCl 2 MgCl 2 Salt..
Per cent sucrose inverted
33.80
35.46
39.68
40.65
47.60
is
50.01
50.01
The
influence of different salts of the
same base
shown
in the fol-
lowing series, where 50 c.c. of solutions containing 40 per cent sucrose and 10 per cent invert sugar were heated at 100 C. for 2 hours with a quantity of different potassium salts equivalent to 1.75 gms. Cl in KC1.
Salt
K-acetate K-tartrate K-oxalate

(
KC1O 3
3 32
K SO KNO
2
KI KBr KC1
Per cent sucrose

inverted
Q()
Q07
Q 3Q
3 go
4.944.946.276.27
inverting power of the different salts is seen to follow the positions of the metal and acid in the electro-chemical series, the salts of the
The
weakest bases and strongest acids having the highest power of inversion. The substitution of other reducing sugars was found by Geerligs to produce the same
effect as glucose and fructose in increasing the inverting power of neutral salts. Non-reducing sugars, such as rafnnose,
had no
sensible action.
action of reducing sugars in increasing the inverting power of has been explained by the formation of basic sugar compounds, ions being thus facilitated. the hydrolysis of the salt and formation of
salts
The
MA +
Salt
C6 H O6
12
Glucose
+ HOH Water
MOH
Hi 2
+ H
A.
Basic glucose
Ionized acid.
compound
Deerr,* who has recently made a study of the question, concludes that the combined influence of glucose and neutral salts does not produce inversion. This conclusion, which is exactly opposite to that of
Geerligs, leaves the subject
inverting power, which different salts under the varying conditions of manufacture
open to further investigation. may have upon sucrose, and analysis, is a factor which the chemist must always bear in mind.
The
INVERSION OP SUCROSE BY INVERTASE
Occurrence of Invertase. The most important inverting agent of sucrose from a physiological point of view is invertase. This enzyme is found widely distributed in the and animal vegetable kingdom, being
*
Bull. 35,
Hawaiian Sugar Planters' Experiment Station.
THE DISACCHARIDES
secreted
669
In-
by
all
living cells
where sucrose undergoes metabolism.
many bacteria, in nearly all yeasts, in different as moulds, Aspergillus and Penidllium, and in the leaves, buds, fruit, reserve organs and other tissues of many higher plants, where sucrose
vertase
is
occurs in
utilized either for the building
up
of
new
tissue or for transportation
to points of growth.
In the animal kingdom invertase
is
found in the intestinal juice and
other fluids of the body. Extracts prepared from the mucous membrane of the intestines, from the kidneys and other organs are strongly
inverting.
sects; its
to.
found in the digestive tract of many inhoney sac of the bee has already been referred While the invertases from different sources resemble one another
Invertase
is
also
presence in the
in their hydrolytic action in behavior. It

is
upon
sucrose, they
show
certain differences
supposed, therefore, that the inverting enzymes constitute a group, the different members of which are not strictly identical. On account of the difficulty of preparing perfectly pure
preparations of invertase, identity or difference of the
sources.
it
has been impossible to determine the enzyme from the various plant and animal
Invertase
is
Preparation of Invertase.
this source.
best obtained from yeast,
and various methods have been devised
Some
for preparing the enzyme from authorities recommend mixing fresh washed yeast
with powdered glass or sand and air drying. The mass is then ground in a mill or mortar and extracted with cold water using a powerful
press to increase the extraction. A more active preparation of invertase than that obtained
above process in which yeast

is
is
obtained by the method of O'Sullivan and
by the Tompson*
subjected to autolytic digestion. Pure fresh brewer's drained and then set aside in a covered jar for several yeast washed, weeks at ordinary temperature until the mass has liquefied. A dark
is
yellow solution
II Fischer,
is
obtained which can be purified and decolorized by bone black. The autolysis may be hastened by first filtering through the life cell of the with chloroform as recommended by destroying yeast
is
The method
as follows:
of
Hudson J
for preparing a stock solution of invertase
Break up 5 pounds of pressed yeast, which may be either baker's or brewer's yeast, add 30 c.c. of chloroform to it in a closed flask and allow it to stand at room temperature (20 C.) over night. By the morning, the solid mass will have become fluid and it
*
J.
"
Chem.
Soc., 57, 834-931.

t J,
t Ber., 27,
2,
2985.
Ind. Eng. Chem.,
143.
670
SUGAR ANALYSIS
should then be filtered through filter paper, allowing several hours To the filtrate add neutral lead acetate until no further for draining. and again filter. Precipitate the excess of lead from forms precipitate
the filtrate with potassium oxalate and filter. To this filtrate add 25 c.c. of toluene and dialyze the mixture in a pig's bladder for 2 or 3 days
against running tap water.
The
dialyzed solution
is
colorless, per-
fectly clear after filtration, neutral to litmus, has a solid content of about one-half of one per cent, an ash content of a few hundredths of one per cent, will keep indefinitely in an ice box if a little toluene is kept on
its
surface to prevent the growth of microorganisms,
and
is
exceed-
The invertase solution does not ingly active in inverting cane sugar. reduce Fehling's solution." The solution of invertase prepared by this
method
gives a dextrorotation of 1
is
V. in a 400-mm. tube.
Invertase
precipitated from solution
by adding about 3
vols. of
strong alcohol. The precipitate is filtered off, and finally dried in a vacuum over concentrated sulphuric acid. The product can be purified
by
redissolving in water
and again precipitating by means of alcohol;

is
such purification, however,

power.
Properties
of
always attended by
loss in inverting
invertase consists of a white water with formation of a yellowish Unless previously dialyzed the product contains neutral solution. considerable mineral matter, the quantity sometimes exceeding 20 per The chemical composition of invertase is not fully known. cent.
Invertase.
Dry
powdery substance
easily soluble in
Barth* found
for an ash-free preparation 43.9 per cent C, 8.40 per and 0.63 per cent S. Osbornef found 44.54 cent H, 6.00 per cent and 6.1 per cent N. The high percentage per cent C, 6.52 per cent of nitrogen, the positive reaction with Millon's reagent and the biuret
test indicate the presence of
an albuminoid group.
Carbohydrates, con-
sisting probably of mannan and pentosan groups, have also been found in invertase. It is uncertain whether these carbohydrate groups are a
constituent part of the

of
enzyme
or like the mineral matter consist only
accompanying
impurities.'
Conditions Affecting the Activity of Invertase. The inversion of invertase molecule of water consists in the addition of one by to each molecule of sugar, but the mechanism of this process is not as
sucrose
yet understood.
It is supposed by some that the configuration of the must conform in certain respects to that of the sugar hydrolyzed enzyme and this is used as an argument for the presence of a carbohydrate
group in invertase.
*
Fischer has likened the relation of

t
enzyme
to
Ber., 11, 474.
Chem. News,
79, 277.
THE DISACCHARIDES
671
sugar to that existing between a key and lock, the shape of the key permitting it to unfasten only that lock to whose structure it corresponds.
The
action of invertase being purely catalytic, a small
amount
of
enzyme can invert almost unlimited quantities of sucrose. O'Sullivan and Tompson found in fact that a preparation of invertase, which had already inverted 100,000 parts of sucrose, had lost none of its activity. To secure Influence of Acids and Alkalies on Activity of Invertase. the maximum inverting power invertase must be allowed to act in a weakly acid solution. The acidity for acids, which are largely dissociated as hydrochloric
acid, should not
greatly exceed
ft/ 1000.
An
For acidity much above n/100 HC1 will completely destroy invertase. acids which are only slightly dissociated, as acetic, the acidity may exceed 100 times the concentration permissible for hydrochloric acid. In analytical work it is best to use invertase in an acetic acid solution;
an
acetic acidity just sufficient to redden litmus
was found by
Hudson*
alkali; in
to give the best results.
is rendered completely inactive by small amounts of such cases the original activity may be regained by restorAddition of alkalies in large amount ing the proper degree of acidity. destroys the enzyme completely.
Invertase
by means
The inversion velocity of sucrose Rate of Inversion by Invertase. of invertase has been a subject of considerable study and the
conclusion of early observers has been that the inversion does not follow the formula for a unimolecular reaction, such as is obtained by inversion with acids. O'Sullivan and Tompson,f however, showed, in 1890, that in following the inversion with invertase a serious error existed in the polarimetric reading if the mutarotation of the freshly liberated sugar was not considered. To quote from these authors: "The dextrose formed by the action of invertase on cane sugar is initially in the birotary state, and, therefore, the optical activity of a solution undergoing inversion is no guide to the amount of inversion that has taken place. If a caustic alkali be added to a solution undergoing inversion, and the optical activity be allowed sufficient time to
become constant, it is a true indicator of the amount of inversion that had taken place at the moment of adding the alkali." When the error due to mutarotation is thus corrected, the inversion by invertase was found by O'Sullivan and Tompson to follow the same
unimolecular formula as by inversion with acids. The action of invertase upon sucrose has recently been studied by
*
J.
t J.
Ind. Eng. Chem., 2, 143. Chem. Soc., 57, 927.
672
SUGAR ANALYSIS
*
and the conclusions of O'Sullivan and Tompson were fully confirmed. Hudson, for example, found for the apparent and real inversion rate of by invertase the following values:
Hudson
TABLE XCVI
Apparent and Real Rates of Inversion of Sucrose by Invertase
Time
(t).
THE DISACCHARIDES
of
673
about about
+109
17,
and the
freshiy liberated or a-fructose a rotation of
the combination of these values, when the a-glucose and a-fructose are molecularly united, giving the specific rotation of sucrose,
i.e.,
[a] D
sucrose
(109 X =^
180)
(17
180)
The Influence of Concentration of Invertase on Rate of Inversion. velocity of inversion with invertase was found by O'Sullivan and Tompson to be proportional to the concentration of enzyme. This
proportionality was tested by Hudson solutions of varying concentration.
and found to be true
for sucrose
following table by Hudson shows the percentage inversion of three sucrose solutions using different concentrations of invertase for different periods of time. In making
The
the experiments small quantities of invertase solution were diluted to |, J, J and |, and 10 c.c. of these dilutions added to 100 c.c. of stock
solutions of sucrose, the concentration of the resulting solutions being 45.5 gms., 90.9 gms. and 273 gms. sucrose per liter.
TABLE XCVII
Influence of Concentration of Invertase on the Rate of Inversion at 30 C.
674
SUGAR ANALYSIS
The Influence of Concentration of Sucrose on Activity of Invertase. the concentration of suinfluenced by activity of invertase is greatly
crose.
This
is
shown
in the preceding table
by Hudson from which the
following figures are taken:
THE DISACCHARIDES
TABLE XCVIII
Rate- of Destruction of Invertase by Alcohol
Concentration of
alcohol
675
(volume per cent)
676
SUGAR ANALYSIS
others, who have investigated this phenomenon, show that in presence of sucrose invertase can withstand higher temperatures and higher con-
The action of centrations of alcohol than where no sucrose is present. fructose in protecting invertase from destruction by acids, alkalies and hot water is shown in Table by Hudson and Paine* where
XCIX
the rates of destruction are expressed as per cent of the rate for the destroying agent when no fructose is present.
The property which sugars have of protecting invertase from destruction has been noted in case of other enzymes (as diastase); the phenomenon can be explained by assuming that the invertase forms a
combination with the sugar which
pure enzyme.
is
less easily
destroyed than the
COMPOUNDS OF SUCROSE
Owing to the absence of free aldehyde or ketone groups sucrose does not form hydrazones, osazones, oximes or other compounds such as are
characteristic of the reducing sugars. Acetic anhydride under varying conditions gives a number of acetates, and benzoyl chloride in presence of sodium hydroxide gives several benzoates of sucrose. These com-
pounds have, however, but

passed over.
little
importance and their description
is
The most important compounds of sucrose from the analytical and technical standpoint are the saccharates, or sucrates, which are formed by the combination of sucrose with various metallic bases.
Saccharates of the Alkalies.
By
treating alcoholic sucrose solu-
tions with concentrated potassium or sodium hydroxide, gelatinous saccharates are precipitated of the formulae Ci 2 2 iKOn and Ci 2 2 iNaOn.
The compounds
are soluble in water
in strong alcohol.
The
dilute alcohol, but insoluble alkali monosaccharates are also formed in
and
aqueous solutions of sucrose after addition of potassium or sodium hyDubrunfaut in fact noted that after droxide, even in slight amounts.
addition of sodium hydroxide to sucrose in equal molecular proportions the specific rotation sank to a fixed value, further addition of alkali pro-
ducing no change.
ing to
The
specific rotation of
:
sodium saccharate accord0.00039944 q 2
Thomsenf
[a] D
follows the equation
= + 56.84 + 0.011359 q +
in
which q is the per cent water in solution. The depressing influence of sodium hydroxide and potassium hydroxide upon the rotation of sucrose, through formation of saccharates, may introduce an error in
*
J.
Am. Chem.
Soc., 32, 988.
t Ber., 14, 1647.
THE DISACCHARIDES
certain polarimetric
677
free alkali is first neu-
measurements unless the
tralized (preferably by means of acetic acid). Saccharates of the Alkaline Earths.
The most important
sac-
charates from the technical standpoint are those of the alkaline earths. In the formation of these the sucrose molecule can combine with one
or
more molecules
of the base.
In case of calcium there are three well

bi-
characterized sucrose
tetra-,
compounds the mono-,
and trisaccharates;
hexa- and octosaccharates have also been described. The structural constitution of these and other saccharates is not as yet understood, the place and manner of attachment of the base to the
sucrose molecule not having been established. It is supposed that the bivalent metals are attached to the sucrose molecule by only one valency, as, for example, Ci2
rate.
H iOn
2
Ca
OH
in calcium
monosacchais
The
existence of sucro-carbonates in which the bivalent metal

is
united both with sucrose and the carbonic acid radical

this supposition.
explained upon
Calcium monosaccharate
finely
powdered quick lime
low temperature.
formed by dissolving sucrose and fresh water at The compound is then precipitated from solution by
is
in equal molecular proportions in
strong alcohol; as thus prepared
it
has the formula:

2
Ci 2
H O n CaO + 2 H O,
22
-
the water of crystallization being expelled by drying at 100 C. Calcium monosaccharate consists of a white amorphous substance, easily soluble in cold water but insoluble in strong alcohol; its aqueous
solutions upon warming become turbid, but the turbidity disappears on recooling. Upon heating its solutions calcium monosaccharate is decomposed into calcium trisaccharate and free sucrose.
CuHzaOn CaO = Ci 2 H 22 On
-
CaO
2Ci 2 H 22 O n
Sucrose.
Calcium monosaccharate
Calcium trisaccharate
Calcium bisaccharate
is
best prepared, according to Lippmann,*
by
adding fresh finely powdered quick lime, free from hydroxide, to a cold aqueous solution of sucrose using 2 molecular parts of CaO to 1 of
cooling the solution with ice beautiful white crystals 2 CaO. 22 On Crystallization at the bisaccharate, and with takes difficulty, higher temperatures place which is then obtained, contains water of crystallization. Calcium
Ci 2
22
On.
Upon
will
separate of the composition Ci 2
bisaccharate
the solution
is
it is
soluble in about 33 parts of cold water; upon boiling decomposed into the trisaccharate and free sucrose.
-
3 Ci 2
22
On
2
*
CaO =
2 Ci 2
22
On
CaO
Ci2
22
On.
Calcium bisaccharate
Sucrose.
678
SUGAR ANALYSIS
Calcium trisaccharate is formed upon boiling solutions of the monoand bisaccharate as above described. It is also produced as a granular precipitate by adding fresh finely pulverized quick lime to an alcoholic
solution of sucrose using 3 molecular parts of

2
CaO
to
of
obtained, after drying over concentrated 4 2 O, one molesulphuric acid, has the formula Ci 2 22 On 3 CaO cule of water, however, being given off in vacuo. The trisaccharate as
Ci 2
H 2On;
the
compound thus
+ H
prepared from hot aqueous solutions has 3 molecules of water. Calcium trisaccharate is a white granular compound, soluble in 100 parts of cold and in 200 parts of hot water.
is
employed technically
in the separation of
sucrose from beet molasses.
In the old elution process of Scheibler*
the molasses was mixed with an excess of freshly burned, finely powdered quick lime, and the porous mass of saccharate thus obtained freed from The elution method is supimpurities by washing with dilute alcohol.
* which planted at present by the trisaccharate process of Steffen carried out as follows. The molasses after diluting to 12 to 14 Brix
is
is
treated in the cold with freshly burned quick lime, reduced to the fineness of dust, in the ratio of 80 to 150 parts by weight of CaO to 100 of
sucrose. Constant agitation of the solution is necessary in order to secure proper distribution of the lime and to prevent too great an increase in temperature. The granular precipitate of trisaccharate is filtered cold through filter presses, washed with cold water and then
either used for saturating the diffusion juice, or worked up separately for sucrose by decomposing with carbon dioxide in aqueous suspension.
CwHaOii
CaO
+
in
3C0
2
Ci 2
H On
22
+
CaO
CaCO
3.
Carbon dioxide
Sucrose
Calcium carbonate.
Double saccharates,
rate
ates
is
which one molecule of
in the trisaccha-
replaced
by
K O or Na O, have also been formed.

2
Sucro-carbon-
have also been prepared; the exact nature of the latter, to which such formulae as Ci2 H 22 On 6 CaO 3 C0 2 have been given, is unknown.
Strontium monosaccharate is best obtained according to Scheiblerf by treating a 20 to 25 per cent solution of sucrose at 70 to 75 C. with equal molecular parts of crystallized strontium hydroxide (Sr(OH) 2 8 2 O)
+ H
and allowing the supersaturated solution to cool with exclusion of the carbon dioxide of the air. By adding a few crystals of monosaccharate from another preparation and agitating the solution, strontium monoFor a very complete description of the osmose, elution, strontia and other processes for desaccharifying molasses see Ware's " Beet Sugar Manufacture and " Refining (1907), Vol. II, 466-510, or the works of Claassen, Newlands, Rumpler,
*
Stohmann and
t Her., 16,
others.
984.
THE DISACCHARIDES
679
saccharate will separate out in cauliflower-like masses of white crystals with a composition corresponding to the formula Ci 2 22 Ou SrO 5 2 0.
+ H
The compound
of crystals.
dissolves in
supersaturated solutions,
warm water with great readiness to form which may be cooled again without separation
its
Upon
is
heating
solutions above 60
C. strontium
mono-
saccharate
decomposed
into bisaccharate
and
free sucrose.
2 Ci 2
22 Oii
SrO
Ci 2
22 Oii
2 SrO
Ci 2
22
On.
Strontium monosaccharate
Strontium bisaccharate
Sucrose.
is best prepared according to Scheibler* by strontium hydroxide in a boiling 15 per cent dissolving crystallized As soon as the molecular proportion of strontium to sucrose solution.
Strontium bisaccharate
1 the bisaccharate begins to separate. When the molecular proportion of strontium to sucrose exceeds 3 1 the separation of sucrose as strontium bisaccharate is almost quantitative after 8 to 10
sucrose exceeds 2
minutes' boiling. Strontium bisaccharate consists of white granular The compound is soluble crystals of the formula Ci2 22 On 2 SrO.
about 84 parts of boiling water but is insoluble in alcohol and in For the complete precipitation of strongly alkaline aqueous solutions. sucrose as bisaccharate the third molecule of strontium hydroxide can, therefore, be replaced by other alkalies such as sodium or potassium
in
hydroxide.
When
strontium bisaccharate
is
mixed with cold water
it
is
de-
composed, there being obtained a solution of the monosaccharate and free strontium hydroxide, the latter separating out in the crystalline
form.
Ci 2
If
22
On
2 SrO
+H
Ci 2
22
On SrO
-
Sr(OH) 2
dioxide the monosaccharate
the filtrate from the strontium hydroxide be saturated with carbon is decomposed into sucrose and strontium
carbonate.
By evaporating the clear filtered solution, the sucrose is recovered in the crystalline form. The method of precipitating sucrose as strontium bisaccharate is employed analytically for detecting sucrose in plant materials (p. 647)
;
it
also constitutes the basis of the strontium process for recovering
sucrose from beet molasses.

diluted molasses
and strontium hydroxide
In the Scheibler t strontium process the (2J to 3 molecules of stron-
tium to 1 of sucrose) are heated to 100 C. with constant agitation for about 30 minutes. The precipitated bisaccharate is then filtered off and washed hot with 10 per cent strontium hydroxide solution, until the
* t
Deut. Zuckerind., 31, 867. " Ware's " Beet Sugar Manufacture and Refining (1907), Vol.
Z. Ver.
II,
502.
680
soluble impurities are
SUGAR ANALYSIS
removed and the precipitate
then cooled for
1 to
is nearly white. The 2 days at a temperature of 5 to 10 C., when it decomposes, according to the preceding equation, into a solution of the monosaccharate and crystallized strontium
washed bisaccharate
is
hydroxide.
The
latter is separated
by
centrifugals
monosaccharate carbonated.
The
filtrate
and the solution of from the strontium carbonate
(which is reconverted into strontium hydroxide) is a sucrose solution of about 97 per cent purity, and can be worked up directly into white The strontium bisaccharate process at the present time is largely sugar. replaced by the Steffens calcium trisaccharate method.
Barium monosaccharate is obtained by warming 100 parts of a 6 per cent aqueous sucrose solution with 20 parts of a 20 per cent barium hydroxide solution and allowing to cool unexposed to the carbon dioxide The compound may be prepared more easily by employing of the air.
aqueous solutions of sucrose. Barium monosaca white crystalline compound with a composition correspondIt is soluble in 47.6 parts of water ing to the formula Ci 2 H 22 Oii BaO. at 15 C., easily soluble in aqueous sucrose solutions, but insoluble in
alcoholic instead of
charate
is
alcohol or in aqueous barium hydroxide solutions. The compound is in contact with water carbon but the last traces decomposed by dioxide,
difficulty; to facilitate the sepathe solution after ration, carbonating may be heated with gypsum or ammonium sulphate, the traces of barium remaining in solution being
of
barium are precipitated only with
precipitated as sulphate.
On account of
barium
tries.
for separating sucrose
the poisonous character of some of its salts, the use of from molasses is forbidden in many coun-
In Italy,* however, the barium saccharate method has proved and is still employed on a large scale, no injurious effects seeming to attend the use of the sugar thus prepared. In the Italian
successful
Be.,
process the barium hydroxide solution is made up at 38 to 40 degrees and the molasses of 38 to 42 degrees Be*, added at a tempera-
to 50 C. The mixture is rapidly stirred and the barium monosaccharate, which soon becomes granular, allowed to settle. With normal molasses the barium hydroxide is used in the proportion of 1 molecule for each molecule of sucrose, plus an extra TV molecule for
ture of 45
The monosaccharate is then washed, decomposed aqueous suspension with carbon dioxide and the filtrate from the barium carbonate evaporated to crystallization. The yield of sugar by the process is about one-third the weight of beet molasses. In both the barium and strontium saccharate processes the barium
the non-sugars.
in
*
Viewegh, Z. Zuckerind., Bohmen, 34, 38.
THE DISACCHARIDES
and strontium are recovered and worked up again
continued use.
Miscellaneous Metallic Compounds of Sucrose.
681
into hydroxides for
In addition to the
saccharates of the alkalies and alkaline earths a large number of compounds of sucrose with other metals have been prepared, such as
lead
saccharates of iron, aluminum, chromium, manganese, nickel, copper, and mercury. Some of the saccharates mentioned, as those of iron, are used medicinally. Lead saccharates of the formulae Ci 2 22 On PbO,
Ci H 2On
2 2
PbO and
and these compounds

line sucrose solutions
are described in the literature, 2 22 Oii are sometimes formed in the clarification of alka3
Ci H
PbO
by lead subacetate with introduction of considerwork of analysis. Soluble lead saccharates may affect the polarimetric' reading, and precipitation of insoluble lead saccharates
able errors in the
introduces a loss in the determination of sucrose.
In connection with the formation of soluble saccharates there should be mentioned the property which sucrose has of preventing or retarding the precipitations of iron, aluminum, cobalt, nickel, copper
and other metals from solution by means
ammonium
of sodium, potassium and In such cases metallic-sucrose complexes are hydroxides. The followis not understood. of which constitution the exact formed, have been which of the formulae are given to a few examples ing
2 Ci 2 H 22 On Fe2 O 3
H22 On 5 CuO + Na2 0; from a study of the electric Kahlenberg* that the metals do believes of such of solutions complexes conductivity not exist in the dissociated condition of an ordinary salt solution but
such compounds as have been isolated, Ci 2
+ 2 NaÔ.
form of complex sucrose-metal ions. Characteristic qualitative tests for detecting Tests for Sucrose. small amounts of sucrose in presence of other sugars are wanting. In
in the
sucrose as one of
such cases the only certain means of identification is to precipitate the its saccharates, preferably strontium bisaccharate,
and
to determine the optical
after liberation
from
its
and chemical properties compound by means of carbon
of the
sugar
dioxide.
The
determination of specific rotation or reducing power before and after inversion with hydrochloric acid or invertase is also valuable as a
means
of identification.
Sucrose in presence of inverting agents will
of course give any of the reactions described for d-glucose and d-fructose. The deep violet coloration which even very dilute sucrose solutions give and sulphuric acid is also given by solutions of invert with
a-naphthol
sucrose with an alkasugar. The violet coloration obtained by heating line solution of cobalt nitrate was formerly regarded as a characteristic
*
Z. physik.
Chem.,
17, 616.
682
SUGAR ANALYSIS
test is
reaction; other sugars, however, give similar colorations so that the not reliable. The colorations which sucrose gives with mor-
phine, codeine, aconitine, veratrine and other alkaloids in presence of sulphuric acid is also given by invert sugar; the same is also true of the blue coloration obtained by treating a sucrose solution with ammonium molybdate in presence of sulphuric acid.
Configuration of Sucrose.
A number
of constitutional formulae
have been assigned to sucrose. The following arrangement by Wohl* and Fischer f is the one most generally preferred, although the exact configuration is still a matter of doubt:
CH OH
2
CH OH
2
HOCH
H
d-Glucose radical.
d-Fructose radical.
The above arrangement contains no free aldehyde or ketone group which explains the non-reducing property of sucrose. The cleavage
into d-glucose
and d-fructose by inversion

with a
*.
is
supposed to take place at

as yet
the
O atom marked
The
synthesis of sucrose from glucose
and fructose has not
been accomplished.
MALTOSE.
Maltobiose.
Malt
sugar.
Cerealose.
The formation
extract
later
of a hitherto unknown sugar by the action of malt upon starch was noted by De Saussure J in 1819; some years Dubrunfaut made a further study of the sugar and gave it the
name
maltose.
one of the most widely distributed however, that maltose is found in plants almost entirely as a transition, and not as a reserve, carbohydrate renders it difficult to isolate the sugar from ordinary plant substances in In the vegetable kingdom maltose has been observed large amounts. in the leaves of many plants, in young twigs and buds, in yeast, soja
is
Occurrence.
Maltose
fact,
disaccharides.
The
Ber., 23, 2084.
t Ber., 26,
2405.
Ann. chim. phys. [2], 11, 379. Ann. chim. phys. [3], 21, 178.
THE DISACCHARIDES
683
beans, rice and other substances; it is found most abundantly in starchy seeds at the time of germination when it is formed together with
dextrin
by the action
of diastatic
enzymes upon
starch.
The
maltose,
which
thus formed, is itself quickly hydrolyzed by other enzymes (glucases), so that the amount of free maltose occurring at any one time is relatively small. In the animal kingdom maltose has been observed in abnormal urines, in the intestinal tract, in the blood, liver
is
and muscular tissues. Its occurrence in the animal organism is no doubt largely due to the action of amylolytic enzymes upon the starchy
matter of the food.
Diastatic
distributed in
Enzymes.
Diastatic enzymes or amylases are widely both the vegetable and animal kingdoms. Aqueous ex-
tracts of barley, oats, rye, rice and other cereal grains as well as of many seeds; extracts of the blossoms, buds, leaves, roots, etc., of
many
plants, and also of many moulds, bacteria, fungi, lichens, etc., possess the property of converting starch into maltose and dextrin. In the animal kingdom amylases are found in the saliva (ptyalin), in
the pancreatic juice
(pancreatin),
in the
mucous
secretions of the
stomach and
intestines,
and
in the liver, kidneys
their presence has also
been reported in
and other organs; blood serum, in the lymph
and even
in urine
and milk.
The
fresh aqueous extract of

seeds,
many
plant substances, such as starchy
grains and grains and
have relatively but little diastatic power; if such seeds, however, are moistened and allowed to germinate before making the extract, the starch converting power is found to
undergo a marked increase. In such cases the amylase is supposed to be derived from an anterior substance, or zymogen, which is itself inactive.
diastatic
following experiments by Salamon* show the increase in power during the germination of barley. The values are exin terms of Lintner's scale (p. 513) and are calculated in each pressed case to a common basis of 2 per cent moisture.
The
Day.
684
SUGAR ANALYSIS
results
The
show a
20-fold increase in diastatic
13 days of germination, although at certain stages there parent decrease upon succeeding days.
power during the was an ap-
Malt.
studied
The
diastases of germinated barley (malt) are of great
importance in the
more than any of the amylases.
barley is first has absorbed about 50 per cent its weight of moisture. The barley is then allowed to germinate for 9 to 12 days upon a floor in heaps about
1
brewing industry and have for this reason been In the preparation of malt, raw steeped for 2 to 3 days in water at 10 to 13 C. until it
foot in depth.
The heaps
are turned several times each
wooden shovels
in order to secure proper aeration
day with and even distribution
of temperature, the latter being maintained as nearly as .possible at 15 C.; the grain is also sprinkled with water at intervals in order to
maintain proper conditions of moisture. After germination has proceeded to the desired extent, as determined by the growth of the rootlets and acrospire, the fresh malt is transferred to a drying kiln,
where 45 C.
it
is
heated at about 25
to 35
C. for the
first
day, at 40 to
65 temperature varying from 85 to 110 C., according to the character The gradual elevation of temperature is necesof the malt desired. sary, as diastase, like invertase and other enzymes, is extremely sensitive to heat in presence of moisture, although when perfectly dry the enzyme can withstand a much higher temperature. The diastatic
for the second day, at 50 to 55 for the third day and at 60 to The kiln is then gradually raised to a final for the fourth day.
power
of
the
green
malt
process, however, being
is considerably reduced by the drying only one-sixth to one-third of its original
amount. In the process of malting a

in
series of important changes take place the carbohydrates of the grain. In the first place a considerable amount of the conversion products of the starch are consumed by res-
piration, over 10 per cent of
carbon dioxide being given off by the malt during germination. The maltose, which is produced by the action of the amylase upon the starch, is hydrolyzed into glucose by the glucase. Synthetic processes also take place; the reducing sugars absorbed by
the aleurone
turn, as
it
cells
and scutellum are
built
up
into sucrose, the latter, in
contributes to the growth of the plant embryo, being hydrolyzed into glucose and fructose. The following analyses by O'Sullivan* give the per cent of different sugars in two samples of barley before and
after germination.
* J.
Chem.
Soc. (1886), p, 58.
THE DISACCHARIDES
685
686
SUGAR ANALYSIS
1872, was the first to subject the action of malt diastase upon starch to a careful study, and since then a large number of investigators have
made
the question an object of research. Owing to the complexity of the Steps in Diastatic Conversion. starch molecule and the indefinite number of intermediate transition
are formed between starch and maltose, such as amylodextrin, erythrodextrin, achroodextrin, malto dextrin, etc., the conversion of starch is a vastly more complicated reaction than the inIt is generally agreed that malt diastase is a mixversion of sucrose. ture of enzymes; the primary phase of starch conversion, consisting in
products which
cytase;
the formation of soluble starch, is attributed to a liquefying enzyme or the remaining steps of the conversion are assigned to an amylo-
which converts the soluble starch into dextrin, and to a which converts the dextrin into maltose; an amylomaltase which converts soluble starch directly into maltose has also been supposed to exist. The difference in behavior of diastases from different sources is no doubt due in part to variations in amount of the
dextrinase,
dextrinomaltase,
constituent enzymes.
Coworkers. The conversion of under ordinary conditions is not complete, the reaction coming to a resting stage or condition of This is represented according to Brown and Heron,* and equilibrium. Brown and Morris f by the equation:
Theory
of
Brown and
his
starch into maltose
by means
of diastase
10C 12 H20
Starch
10
+ 8H
8C H On +
12
22
12
H O
20
10 .
Maltose
Dextrin.
Brown and
Millar { in a later research
show that the dextrin thus
formed, upon prolonged treatment with diastase, breaks up into glucose as well as maltose, and to explain this and other facts give the equation
:
100 (Ci 2 H2oOio)

Starch
+ 81 H
80 Ci 2 H 22 On
Maltose
(C 6
10
5 )39
12
6.
Dextrin.
In other words 100 parts of starch yield 84.44 per cent of maltose. In this connection it is interesting to note that Sherman and Kendall found with pancreatin a tendency to equilibrium when the weight of maltose reached about 85 per cent of the initial weight of starch.
Starch according to
*
J.
Brown and
Millar
||
has a molecular weight of
t J. }
II
Chem. Soc. Chem. Soc. J. Chem. Soc. J. Am. Chem. J. Chem. Soc.
Trans. (1879), 35, 596. Trans. (1885), 47, 527. Trans. (1899), 75, 333.
Soc., 32, 1087.
Trans. (1899), 75, 333.
THE DISACCHARIDES
687
34,200 and consists of four similar maltan groups, (Ci2H2oOi )2o, and one dextran group, (C 6 Hi 5 )4o, combined in the following arrangement:
hydrolysis with diastase the dextran complex A is split off, the stable dextrin 39 (C 6 Hio05 ) C6 Hi 2 O6 which undergoes no forming further change under the ordinary conditions of conversion. The
Upon
maltan complexes, B, on the other hand, are decomposed at the O linkages which join the Ci 2 -groups and give rise, as the hydrolysis proceeds, to a
series of maltodextrins of diminishing as the final end product.
molecular weight with maltose

of its conversion
The above formula

acceptance.
for starch
and the theory
by
diastase require, however,
much
additional confirmation before final
serves, however, as a good illustration of the involved. are which complex problems Roux. of and According to the recent conTheory Maquenne clusions of Maquenne and Roux * starch is to be regarded not as a com*
The example
Ann. chim. phys.,
9, 179.
688
SUGAR ANALYSIS
pound but as a mixture of amylocellulose, or amylose, and amylopectin. The following conclusions are taken from the work of these authors:
Amylocellulose
is
identical with the granulose or soluble amylose of
previous writers, and constitutes 80 per cent to 85 per cent of natural Under amyloses are comprised those substances which starch grains.
are colored blue
by
iodine, are perfectly soluble in potash solution or
superheated water and are saccharified without producing residual The less condensed amyloses form the different soluble dextrins.
starches; the more highly condensed members are not soluble in the pure state except under pressure, at 150 to 155 C., but they form with
members eutectic mixtures, perfectly soluble in The transformation of a lower amylose into a higher, less
the lower
boiling water. soluble homo-
logue does not appear to take place outside the living cell. Amylose can assume at the same temperature two distinct forms; a soluble form immediately saccharified and colored by iodine, and a solid form which
malt and gives no reaction with iodine. The latter is perhaps a polymeric form of the first. Solutions of amylose give with iodine a coloration about one-fourth more intense than those of natural starch. Starch grains are colored with iodine because a part of its amylose exists as a solid solution. Starch paste may retrograde, owing to the of the fresh paste holds in solution. By which crystallization amylose means of this property the crude amylose may be purified, and obtained in grains which resemble the original starch in microscopic appearance. Besides amylose, natural starch contains 15 to 20 per cent of a mucilaginous substance, amylopectin, which differs from amylose by swelling up without dissolving in boiling water or alkaline solutions, by being only very slowly saccharified by ordinary diastase and by giving no reaction with iodine. Starch paste is simply a perfect solution of amylose,
resists
rendered viscous by amylopectin.

proceeds in
The
saccharification of starch paste
two successive phases, a rapid phase of a few hours and a The saccharification of the slower phase which lasts several days. so-called residual dextrins, makes the The amylose rapid phase. up which accompany maltose in those worts which are imperfectly saccharified, result from the liquefaction and incomplete hydrolysis of the amylopectin. Malt extract is susceptible to auto-excitation as a probable result of the proteolysis of its albuminoids; this excitation is observed at all temperatures at which the amylase may exist undestroyed,
always accompanied by a partial coagulation. Acids stimulateby producing the same condition of equilibrium which results from auto-excitation. Their effect, however, is generally less favorable than that of the latter, since the stability of the amylase
is
and
the activity of malt
THE DISACCHARIDES
is
689
Diastatic saccharification, to obtain a maximum effect, out in an alkaline medium. The optimum is obtained carried be should first neutralizing the paste and then adding to the malt solution by
diminished.
enough sulphuric acid to neutralize from one-third to one-half the alkaThe second or slow linity present, using methyl orange as indicator.
phase of ordinary saccharification corresponds to the hydrolysis of the residual dextrins (amylopectin) by means of a special diastase (dextrinase) formed during the auto-excitation of the malt.
The
following results
by Maquenne and Roux show the action
of
malt extract at 50 C. upon starch paste and upon a solution of amylose:
Time.
690
in
SUGAR ANALYSIS
malt diastase and the complexity of the various reactions are con-
sidered, that the efforts to establish a simple law of mass action for starch conversion, such as that observed for the inversion of sucrose, should have met with failure. The fact that the starches of different
still
vegetable origin have in all probability a different molecular structure further complicates the problem. Influence of Temperature upon Diastatic Conversion. The
for saccharification of starch by malt diastase about 45 C., although the point of maximum conversion may lie considerably above or below 45 C., according to conditions. Diastase solutions undergo a great reduction in activity upon warming above 60 to 65 C.; above 75 C. the saccharifying power is completely de-
optimum temperature
is
stroyed.
The effect of temEffect of Mashing at High and Low Temperature. perature upon the different enzymes of malt extract is variable. Malt extract, which has been heated to 75 C. and which has thus lost its
saccharifying power,
it
still
liquefies starch as strongly as ever, converting
almost quantitatively into dextrin.
The optimum temperature
for
the liquefying and dextrin-forming enzymes of malt extract lies, in fact, between 70 and 75 C. It is evident, therefore, that the yield of maltose and dextrin from starch can be controlled to a considerable extent by the temperature of conversion, and this fact is utilized in
Mashing at 70 C. will produce and hence give a beverage of greater body (solid content), than mashing at 45 C. Mashing at 45 C., on the other hand, will produce more maltose and hence give a beverage of higher alcohol content than mashing at 70 C. The composition of worts by the high and low temperature methods of mashing is given in the following table:*
more
dextrin,
the technical operations of brewing.
Character of Wort.
THE DISACCHARIDES
of
691
power undergo a decrease but glucose begins to be formed as one the products of conversion. Malt extracts whose saccharifying power has been weakened by heating are said to be restricted; the
maximum
yield of glucose (12 per cent of total conversion products) is obtained by malt extracts which have been heated at 68 to 70 C. for 15 to 30 minutes. Ling and Davis explain the phenomena of restriction
by assuming that an alteration has been produced in the enzyme molecule so that glucose becomes the end product of conversion instead of maltose. Prior,* however, explains the facts by assuming
that a glucose-forming enzyme (amyloglucase) exists in malt extract and is more resistant to heat than the amylomaltase.
The presence of glucose in malt sirups was at one time regarded as an evidence of adulteration with commercial dextrose or glucose sirup. Perfectly pure malt sirupsf may contain, however, several per cent of glucose if the diastase of the malt has undergone restriction. A large amount of glucose may also be derived from the malt itself, as shown by the analysis of cold water extracts (see table, page 511).
Influence of Acids, Alkalies, Salts, Etc.,
Upon Amylolytic
Action.
minute amounts accelerates the activity of malt diastase; in larger amounts acids have a marked retarding influence upon the enzyme, the degree of retardation following apparently the same rule noted lor invertase and being proportional to the conof acids in
The addition
centration of hydrogen ions. Alkalies and alkaline-reacting salts are very injurious to the action of malt diastase if present beyond the merest trace. A perfectly neutral
medium
is
believed
by some
to be the
most favorable
for diastatic
action, while others maintain that the reaction should be slightly acid or even faintly alkaline. The explanation of these differences of
opinion
is
probably the same as that given by Sherman and Kendall
for pancreatin (p. 694).
Small amounts of the neutral salts of the alkalies and alkaline earths
(chlorides, sulphates, phosphates, etc.), usually accelerate the activity
malt diastase, frequently to a very marked degree. Calcium and barium chlorides seem, however, to have a retarding influence. Addition of sulphates or of salts of the heavy metals in large amounts check
of
silver nitrate or of
the activity of the enzyme, owing probably to precipitation. Traces of mercuric chloride destroy diastatic action completely. Of organic substances albumin and asparagine seem to favor diastatic action.
Alcohol in slight amounts exerts no appreciable influence;

*
Wochenschr. f. Brauerei, 21, 349 (1904). Long and Rendle, Analyst (1904).
692
in larger quantities,
SUGAR ANALYSIS
however, the activity of the enzyme is reduced, or destruction to Formaldehyde in amounts exceedprecipitation owing marked influence. has a cent 0.005 retarding per ing
.
The
upon
ucts,
diastase
destructive action of heat, acids, alkalies, salts, alcohol, etc., is considerably reduced, if starch or its conversion prod-
substances
maltose and dextrin, are present; the protective action of these is similar to that noted for sucrose and fructose upon in-
vertase (p. 675). As to the action upon starch of Action of Other Amylases. other diastases than those of malt, mention will be made only of takadiastase
and
of the animal amylases ptyalin
and pancreatin.
diastase, has in saccharify-
Takadiastase, the best
been employed in
known example of a fungus Japan for an unknown period of time

is
ing starchy materials for the production of alcoholic beverages.
The
enzyme has been separated by Takamine* and
now
a standard
pharmaceutical preparation for the relief of starch indigestion. The patented process of Takamine for its production is as follows: Wheat bran is steamed and then, after cooling, sown with the spores
mould Aspergillus or y zee. The moist bran is kept at a temperature of about 25 C.; in about 24 hours the spores have germinated and the growth of mycelium becomes visible; after about 48 hours,
of the
when the production of diastase has reached its maximum, further growth of the mould is checked by cooling. The material in this conbran felted together by the threads of mycelium, in Japan, where it is used in the same manner as malt. To prepare the enzyme " taka-koji" is extracted with water, the aqueous extract concentrated at low temperature, and then treated with an excess of alcohol. The takadiastase, which is precipitated, is
dition, consisting of
is
called "taka-koji
"
filtered off,
pressed and carefully dried; the enzyme as thus prepared consists of a white powder, easily soluble in water, and has a very
strong converting power.
Stone and Wright f have
made
of a pharmaceutical preparation of takadiastase at 40
a comparative study of the action C. and of a
laboratory preparation of malt diastase at 60 C. Following the conversion of potato starch it was noted that the takadiastase was more rapid in its action during the initial conversion than malt diastase, there being an almost immediate change from the typical blue of the
starch-iodine
other
compound to the reddish and violet tints. "On the hand the complete conversion of the starch into forms which no
* t J.
Am. Jour. Pharm., 70, No. 3; Am. Chem. Soc., 20, 639.
J.
Soc.
Chem.
Ind., 17,
No.
2.
THE DISACCHARIDES
693
longer gave color reactions with iodine was effected much earlier by the malt diastase." The same results were obtained when the saccharification was followed by studying the decrease in specific rotation and the increase in copper reducing power. The results of the work of Stone and Wright show that for a given
short period (15 minutes to 2 hours) the saccharifying power of the takadiastase was superior to that of the malt-diastase preparation, but that for the complete saccharification of starch, especially in cellular
materials, where the starch granules were retained and not readily brought into solution, the malt diastase was more effective; the cellular residues after 7 hours' digestion with takadiastase at 40 C. still gave the iodine reaction when observed under the microscope, while the residues after 7 hours' digestion with malt diastase at 60 C. gave no such reaction. These results were obtained, however, with only one set of enzyme preparations and under only one set of conditions. With
enzyme preparations, and other conditions of temperature, activation, etc., than were employed by Stone and Wright, different redifferent
sults
would no doubt be obtained.
Ptyalin, the amylase of saliva, plays an important part in the digestion of starchy foods; it occurs most abundantly in the saliva of
herbivorous animals.
Ptyalin can be prepared from saliva by precipiThe tating with alcohol, as described under invertase and diastase. optimum temperature for its action is about 40 C., at which point
starch paste is saccharified almost immediately. Raw starch in the process of mastication is also quickly converted into 80 to 100 per cent
sugar.*
Ptyalin,
similar
to
diastase,
contains
several
enzymes,
In some liquefying enzyme, an amylomaltase, an amyloglucase, etc. cases the product of conversion seems to be almost pure maltose; in
other cases a mixture of maltose, glucose and isomaltose (?). The variability of its action is no doubt due to differences in the amount of the
constituent enzymes. Minimal quantities of acid (under 0.002 normal) accelerate the action of ptyalin; large amounts of acid have a retarding influence. Alkalies and alkaline reacting salts are depressing in their
The chlorides, sulphates, etc., of the alkalies also retard the of activity ptyalin if present in large amounts. Pancreatin, the amylase of the pancreatic juice, has recently been
action.
subjected to a careful study by Sherman f and his co workers and the following facts are cited from their work.
Commercial pancreatin, which had been freed from accompanying

* Miiller,
Chem.
t J.
Am. Chem.
Centralbl. (1901), 637. Soc., 32, 1073, 1087; 33, 1195.
694
salts
SUGAR ANALYSIS
by dialysis, was without action upon dialyzed soluble starch. When, however, a neutral salt was added the enzyme was activated as shown in the following experiment: 0.35 mg. pancreatin in 50 c.c. of 1 per cent dialyzed starch at 40 C. for 1 hour showed for various additions of salt the following activities, expressed by weights of reduced cuprous oxide obtained upon heating
with Fehling's solution:
Sodium chloride, mgs. Cuprous oxide, mgs.

results.
0.01
0.1
1.0
10
51
10 87
30
91
60 86
90 85
121
85
Experiments with potassium and
ammonium
chlorides gave similar
of salts are, therefore, not only helpful but are essential to the action of the enzyme.
The presence
influence of acid
Excess of acid or alkali destroys the activity of pancreatin. The and alkalies in minimal amounts is given in the follow-
ing table which shows the action of 0.125 mg. pancreatin upon 0.25 gm. soluble starch at 40 C., sufficient NaCl being added to activate the
enzyme.
Results are given as milligrams of reduced cuprous oxide.
TABLE C
Conversion of Starch by Pancreatin (Effect of Added Acid and Alkali on Solutions Containing Neutral Electrolyte)
THE DISACCHARIDES
starch
695
tion of starch, the effect at first being to accelerate, and then, as the is changed, to retard the speed of saccharification. For a short
period of time an alkaline (and for a long period of time an acid) reaction gives the maximum yield of maltose. This is no doubt one explanation for the variable conditions reported by different investigators
for the
optimum conversion
of starch
by pancreatin and other amylases.
o
03
w
l80
0180
a 2120
100
s
80
20
10
40
50
60 Minutes.
70
'JO
100
110
120
Time curves showing effect of concentration of soluble starch upon the Fig. 199. rate of conversion by pancreatin. .A, curve for 0.5 per cent; B, curve for 2.0 per cent; and C, curve for 4.0 per cent starch solution. (Sherman and Kendall.)
The
effect of concentration of starch
upon the
rate of conversion
by
pancreatin is shown in Fig. 199. A constant quantity of enzyme was allowed to act upon starch solutions of 0.5 per cent, 2.0 per cent and 4.0 per cent strength. It is seen that the initial speed of conversion for a constant amount
of
the same for the different concentrations, but that this speed diminishes more rapidly the smaller the initial concentration of starch. With increasing concentration of starch the time curves ap-
enzyme
is
proach a straight
line.
The
effect of
temperature upon the activity of pancreatin
is
shown
696
in the following table:
SUGAR ANALYSIS
0.5
mg.
of
enzyme was allowed
to act
100
c.c.
of starch solution for 1 hour, in presence of a sufficient
upon amount
of activating salts.
Temperature.
THE DISACCHARIDES
CONVERSION OF STARCH BY ACIDS
697
When
starch
is
heated with acids
it is
converted into glucose ac-
cording to the equation:
(C 6 HioO B )n
Starch
+ nH O
2
= nC H O
6
12
6.
d-Glucose.
With strong
acids the conversion

it is
may
be made in dilute solution at
necessary to employ a higher concentration of acid and, in certain cases, to conduct the hydrolysis under pressure at temperatures considerably above 100 C. in order to secure
100 C.; with weak acids
complete conversion into glucose. While the acid conversion of starch in its final phase proceeds very closely according to the above equation, the different stages of the conversion, as starch to dextrin, dextrin to maltose, maltose to glucose
present the same complexities and uncertainties observed in the conversion by diastase. The best recogFormation of Maltose During Acid Conversion. of starch nized products of the incomplete conversion by acid are gluetc.,
cose
and
dextrin.
The occurrence
of maltose
among
the products of
incomplete acid conversion has been a subject of much dispute; many chemists hold that, while maltose exists as an intermediate product in
the conversion, it is hydrolyzed into glucose almost as quickly as formed, and that the apparent values found for the specific rotation and reducing power of maltose are in reality only the values for mixtures of glu-
and dextrin. Maltose has been separated, however, as its osazone by Rolfe and Haddock* from the acid conversion products of starch and its presence has also been recognized by Sieben,t Vogel,| Weber and MacPherson and other investigators. A great differFormation of Dextrins and Reversion Products. ence of opinion also exists as to the nature of the dextrins which are
cose
formed during the acid conversion of starch. Some chemists believe that only one dextrin (of about [a] D 200) is formed; other chemists, of dextrins having different series a exists there hold that however, rotations and reducing powers and resembling the amylo-, erythro-,
achroo- and maltodextrins of diastatic conversion.

is
An
additional
the formation of reversion products, especially when complication a part of the the starch is hydrolyzed by more concentrated acid, different synthetic and isomaltose form to recombined glucose being
dextrins.
*
Am. Chem. Soc., 25, 1015. t Z. Ver. Deut. Zuckerind, 34, 837.
J.
Chem.
J.
Ztg., 19, 408. Soc., 17, 312.
Am. Chem.
698
SUGAR ANALYSIS
of
Manufacture
Commercial Glucose and Dextrose.

is
The
acid
conversion of starch
of great technical importance, being used in the
manufacture of starch sirups (commercial glucose) and commercial In the manufacture of glucose sirups starch dextrose or grape sugar. (usually corn starch) is mixed with water to a cream of about 20 degrees
Be. and then heated with about 0.06 per cent its weight of hydrochloric acid in copper converters under a pressure of about 30 pounds. The con-
The is controlled by iodine tests and requires about 1 hour. which has a density of about 18 degrees Be., is then neutralized with sodium carbonate, filtered through bone black and evaporated in vacuum pans to the required density, which varies between 41 and 45 In some factories degrees Be. according to the demands of the trade. sulphuric acid is used as the hydrolyzing agent, in which case calcium
version
liquid,
carbonate
is
used for neutralizing.
In the manufacture of dextrose or grape sugar, a much larger amount of acid is used for conversion, frequently 1 per cent or more
of the weight of starch,
is
and the heating is continued until all dextrin hydrolyzed, the end point being indicated by the absence of a precipiThe tate when a little of the solution is poured into strong alcohol.
liquid
vacuum
then neutralized, filtered through bone black, evaporated in to a thick sirup and poured into pans or moulds where it is " " seeded or allowed to solidify; the contents of the pans are usually
is
primed with a
tion.
little
crystallized dextrose to hasten the crystalliza-
The progression of the Upon the Acid Conversion of Starch. hydrolysis of starch by means of acid is described by Rolfe as follows: " The gradual disintegration of the starch molecule and the different stages of the hydrolysis of the products of this disintegration all
Rolfe
go on at the same time, so that the final products of hydrolysis are always present in very small quantity even at the initial stages of the The progression of the hydrolysis manifests itself in the hydrolysis.
The starch paste gradually loses its colloidal nature and passes over to a thin sirup, its viscosity continually deThe dissolved carbohydrate increases in weight but the creasing.
following characteristics:
density effect of a given weight of carbohydrate in a given volume of solution continually decreases. The specific rotation of the carbohydrate, taken as a whole, likewise decreases, while its cupric-reducing
*
Rolfe,
Defren,
"An
Chem.Soc.
"The Polariscope" (1905), p. 175. See also the paper by Rolfe and Analytical Investigation of the Hydrolysis of Starch by Acids." J. Am. 18, 869.
THE DISACCHARIDES
power
699
increases, these values progressively approaching those for dextrose. " The iodine tests are also characteristic; a few drops of iodine solution giving, with the hydrolyzed solutions, at ordinary temperature,
colors
which change progressively as the hydrolysis proceeds from the
deep sapphire blue of the unchanged starch, first to violet and reddish purple, then to a rose madder, and then to a reddish brown, growing lighter as the conversion proceeds, till at a later stage, but before hydrolysis is complete, the iodine gives no color reaction." Maltose is best prepared by the followPreparation of Maltose.
ing method of Herzfeld;* 500 gms. of starch are stirred into 500 c.c. of water at 30 C., 4 liters of boiling water are then added and the paste which is formed cooled to 60 C. Malt extract, prepared by digesting 100 gms. of finely ground malt with 500 c.c. of water at 30 to 40 C., is then added and the liquid kept at 60 C. for 2 hours. The solution is then filtered, evaporated to 750 c.c. and 87 per cent alcohol added until the alcoholic strength of the solution is between 60 and 70 per cent. After standing 24 hours in a closed vessel, the alcoholic solution is decanted from the precipitated dextrin; the alcohol is distilled and
the solution evaporated to a thin sirup. The latter is then extracted with 1 liter of 87 to 90 per cent alcohol, by boiling with successive
The combined extracts, containing portions under a reflux condenser. the maltose, are set aside in a closed flask for 24 hours, filtered from deposited impurities, evaporated to a sirup and then allowed to stand
an open dish at 20 to 25 C. After several weeks' standing the maltose will crystallize either in white concretions or as fine microscopic If the sirup be spread in a thin layer, and then primed with a needles.
in
few crystals of maltose, and stirred at frequent intervals, crystallization will be complete in about 8 days. The crystalline mass is then rubbed to a paste with cold methyl alcohol, pressed between filter paper and recrystallized from hot methyl alcohol, using bone black. Maltose as ordinarily prepared is Properties of Maltose. obtained as the monohydrate C^HÔn H2 0, consisting of fine prismatic needles, which melt upon rapid heating at about 100 C. The water of crystallization is removed only with great difficulty. Upon heating in the air at 100 to 110 C., the water is slowly evolved, but with decomposition of the sugar. If the monohydrate is first dried over concentrated sulphuric acid and then slowly heated up to 90 C. over a strong dehydrating agent (as phosphorus pentoxide), under the vacuum of a mercury pump, the last traces of water are finally removed. The
Neue
Z. Riibenzuckerind, 3, 150; Ann., 220, 200.
700
SUGAR ANALYSIS
anhydrous maltose, as thus prepared, consists of a white amorphous mass and is extremely hygroscopic, absorbing moisture upon exposure to the air with the same avidity as calcium chloride.
Specific Rotation.
The values given

136 to
in the literature for [a] D of
150, the extreme figures being due no doubt to impure preparations of sugar contaminated with water of hydration or with higher rotating dextrins. The values for carefully
maltose range from
crystallized 137 to
and dehydrated preparations
of maltose range
139, the variations in this instance being
from about due to the influ-
mean
tion
ence of temperature and concentration. For ordinary purposes the value +138 may be used. The general equation for concentra-
and temperature
is
The
specific rotation of
given on page 181. maltose hydrate
is
95 per cent of that for the
anhydride.
Freshly prepared solutions of maltose exhibit mutarotation, the

initial rotation,
the
however, as Dubrunfaut first observed, being less than constant value. Parcus and Tollens* found for 1.9074 gms.
8 minutes after solution 15 minutes after solution 30 minutes after solution 1 hour after solution 2 hours after solution 5 hours after solution 24 hours after solution
of maltose anhydride dissolved to 20 c.c. the following values:
'
+ + +
-j-j-j-
119.36 121.01 123.35 128.07 132.97 136.52 136.96
to 20 c.c.
Schulze and Tollens f noted for 2 gms. of maltose hydrate dissolved an initial rotation of 129.42. 95.83 and a constant value of
An
destroys the mutarotation and gives the constant value within a few minutes. As first shown by Brown
addition of a trace of
ammonia
and Morris { maltose at the moment of its formation from starch by means of diastase exists in the low rotating form. Reactions of Maltose. Maltose reduces Fehling's solution about 60 per cent as strongly as d-glucose. If after the end of the reduction the solution is acidified with hydrochloric acid and then again boiled
with Fehling's solution, a second quantity of copper is reduced, in about half the original amount. Maltose is distinguished from the simple
reducing sugars by
tion (p. 336).
its failure
to reduce Barfoed's copper acetate solu-
Oxidation of Maltose. By the action of bromine in aqueous solution maltose is oxidized to maltobionic acid. This was obtained by Fischer and Meyer as a sirup, which upon boiling with 5 per cent sulphuric
*
Ann., 257, 173. Ann., 271, 219.
Chem. News,
Ber., 22, 1941.
71, 123.
THE DISACCHARIDES
acid
701
was hydrolyze'd into d-glucose and d-gluconic acid. This reaction and the reducing properties of maltose indicate that one of the
glucose radicals of maltose has its aldehyde group in the free reactive The oxidation to maltobionic acid is shown as follows: condition.
H oO CH-0-C5H oO4 CHO + O =

1
CsHioOsCH-O-CsHmCX
d-Glucose
radical
COOH
d-Glucose
radical
d-Glucose
radical
d-Gluconic acid
radical
Maltose
Maltobionic acid
Oxidation of maltose with nitric acid gives d-saccharic acid. Action of Alkalies. Maltose upon heating with dilute alkalies unan almost complete loss of optical activity, the sugar molecule dergoes
undergoing partial hydrolysis and rearrangement* with formation of
d-mannose and other products of unknown composition. Upon warming with concentrated alkalies maltose solutions turn dark brown, the maltose being broken up into lower decomposition products among which lactic acid is the most important. The lactic acid thus formed consists, according to Duclaux,f of a mixture of d- and d, 1lactic acid, and under favorable conditions may equal 50 per cent of
d-glucose,
the original weight of maltose. Action of Acids. Maltose on heating with 2 to 3 per cent hydrochloric or sulphuric acid for several hours upon a boiling water bath is
hydrolyzed into d-glucose.
+HO
2
2
d-Glucose.
Maltose
The hydrolysis proceeds much more slowly than the

crose
inversion of su-
yield of d-glucose is nearly but not absolutely quantitative, being 98 per cent to 99 per cent of the theoretical; the 1 to 2 per cent loss is due to destruction of sugar with formation of levulinic
and the
products, etc. acids, according to Sigmond,J follows Wilhelmy's law for a reaction of the first order, the velocity constant k increasing with concentration and rising temperature. W. A.
acid,
humus substances, reversion The hydrolysis of maltose by
and his coworkers found, however, that the values for k as determined from copper reducing power, show a rapid decrease in the The hydrolyzing power of the different later stages of hydrolysis. acids upon maltose follows the same order observed by Ostwald for
Noyes
the inversion of sucrose.
In so far as the various yeasts, Fermentation of Maltose. moulds and bacteria secrete the enzyme maltase or maltoglucase they
*
f
Rec. trav. Pays-Bas, 14, 156, 203.
t Z.
J.
Chem.
Centralbl. (1894), 169.
Am. Chem.
physik. Chem., 27, 386. Soc., 26, 266.
702
SUGAR ANALYSIS
ferment maltose in the same manner as d-glucose. In case, however, the organism does not form maltoglucase, as, for example, Saccharomyces Marxianus, maltose is not fermented. Maltoglucase is formed
in large
is
amount by many
varieties of yeasts, a classification of
which
given in
Table CII, page 714.
Ordinary beer yeast is especially rich in maltoglucase and ferments maltose with the same ease and rapidity as d-glucose, 100 parts of maltose anhydride, according to Jodlbauer, yielding 51.08 per cent alcohol, 49.04 per cent carbon dioxide, 3.95 per cent succinic acid and
glycerol
a total of 105 per cent, and 0.90 per cent of other products which corresponds to the theoretical yield of d-glucose from maltose.
Maltoglucase.
The preparation
of maltoglucase presents consider-
difficulty than that of invertase owing to the greater resistance of the enzyme towards extraction and its greater sensitiveness towards antiseptic agents. According to Fischer and Lindner* the best is prepared by washing the yeast with water, drying enzyme
ably more
for 3 days at ordinary temperathe dried then yeast in a porcelain mortar and expulverizing ture, its times 20 with weight of water for 20 hours at 33 C. As tracting or toluol f are less injurious than chloroform. antiseptic agents thymol been not isolated as yet in the pure form; its soluhas Maltoglucase
upon an unglazed earthen-ware plate
tions
and preparations are always contaminated by other enzymes,
(invertase, amylase, etc.).
The temperature optimum
for the activity
of maltoglucase, according to Lintner and Krober,J is about 40 C. In addition to yeast different varieties of Mucor, Aspergillus, Mo-
Torula, as well as various Amylomyces, form maltoglucase and ferment maltose with production of alcohol. Maltoglucases are also found in many grains, in malt and in most
nilia,
starchy seeds during germination, in peas, beets, potatoes, in the green many plants and in other vegetable organs; the enzyme occurs mostly associated with amylases. The same association also
leaves of
exists in the
animal kingdom, maltoglucases being found in
saliva, in
pancreatic juice
and
in the secretions of the intestines, liver, etc.
Maltose is fermented by nearly all the lactic and butyric acid organisms in the same manner as d-glucose. The same is also true of most oxidizing fermentations. Citromyces Pfefferianus yields about 50
per cent citric acid from maltose, Bad. oxydans produces acetic acid. Oxalic acid, butyl and other alcohols and ethyl acetate are among the
products of special fermentations.

*
Leuconostoc mesenterioides produces

Ber., 28, 984.
t Ber., 28, 1050.
THE DISACCHARIDES
lactic acid
703
is
from maltose but does not produce dextran as
the case
with sucrose.
Maltose contains a free aldehyde group Compounds of Maltose. and the sugar is consequently much more reactive than sucrose, forming methyl and ethyl maltosides, mercaptals, ureides, etc., in the same manner as the simple reducing sugars. In the same way maltose reacts with phenylhydrazine and its substituted derivatives forming a The most important of large number of hydrazones and osazones. the latter from the analytical standpoint is maltose-phenylosazone, Ci 2 H2o0 9 (N NHCeH 5 ) 2 which is formed by heating maltose solutions with an excess of phenylhydrazine acetate for 1 hour. The osazone,
,
owing to
its solubility
it
cooling, pound after recrystallizing melts
when
in hot water, does not crystallize out until after separates in the form of fine yellow needles the com;
upon rapid heating

is
at 206
C. with
in
decomposition.
cold water,
is
Maltose-phenylosazone
only
slightly
soluble
soluble in 75 parts hot water, in 150 parts hot absolute but is insoluble in ether. It undergoes decomposition upon alcohol, with so that the action of hot solutions boiling water, long heating
the heating is continued too far the melting be reduced to 150 C. Maltosazones of low may when the reaction is carried out with are also obtained melting point too little phenylhydrazine or in too small an amount of water. The
if
must not be prolonged;

point of the osazone
melting point and character of the osazone are also greatly modified by other sugars and especially by the different dextrins of starch conversion.
Maltose forms with acetic anhydride a number of acetates of which the octacetate, CfcHi4(CjHjO)sQii, is the most characteristic;
it
consists of white crystals with bitter taste, melting at 157
to 159
C.,
and giving in benzol solution [0:]^ = +76.54, in chloroform [a] D = +61.01 and in alcohol [a] D = + 60.02. Maltose forms with alkalies and alkaline earths a series of maltosates, none of which, however, has the importance of the corresponding sucrose derivatives. Upon treatment with hydrocyanic acid maltose forms a nitrile which yields after saponification maltose carboxylic acid; the latter consists of a
colorless sirup
acid.
5 10
The
and gives upon hydrolysis d-glucose and a-glucoheptonic reaction is expressed as follows:
6
12
C H 05CH-0-C H 05COOH
v
'
+H
= C
H O + C H O COOH.
12
13
d-Glucose
a-Glucoheptonic acid.
Maltose carboxylic acid
Tests.
of other sugars are lacking.
Characteristic qualitative tests for maltose in presence The osazone reaction is one of the best
704
SUGAR ANALYSIS
of maltosazone affording soluble osazones of other
means of identification, the greater solubility an easy means for its separation from the less
sugars;
the influence of
impurities in modifying
the character of
maltosazone must, however, always be borne in mind. The test has been modified by Grimbert* by treating the impure maltosazone with a little cold aqueous 50 per cent acetone and filtering; the maltosazone
separates from the filtrate in pure crystalline form. The inability of certain yeastsâs Saccharomyces Marxianus and
yeast No. 538 of the Berlin Experimental Brewery, to ferment maltose is another means of separation and identification which may be em-
ployed under certain conditions.
The configuration of maltose has not been Configuration. The following provisional formula sugestablished with certainty. Fischer answers, however, to most of the chemical properties gested by
and reactions
of maltose:
H H OHH H
HOH C-C-C-C-C-C OH H OH
2
I I
I
H H H OH H
- O - C-C-C-C-C-CHO
H OHOHH OH
I I
Maltose has not been synthetized as yet with certainty by purely chemical means. The synthesis, however, seems to have been accomplished by the action of certain enzymes
Synthesis of Maltose.
upon glucose
in concentrated solution.
Croft Hillf was the
first
to
discover the synthetic action of enzymes; Hill observed, when extract of dried yeast, or takadiastase, was placed in concentrated glucose solutions, that
a disaccharide sugar was formed.
This sugar he believed
at first to be maltose, and explained its formation by assuming the action of the enzyme to be reversible. Emmerling,J however, in re-
peating Hill's work, believed the disaccharide to be Fischer's isomaltose, and the same conclusion was also reached by E. F. Armstrong. Hill in a later work, while reaffirming his belief in the formation of some maltose, states that
a different isomeric sugar, which he
calls revertose, is
the
main product
of the condensation.
Armstrong Upon Enzymic Synthesis. By action of the enzyme emulsin upon d-glucose for a long period of time Armstrong observed the formation of a disaccharide which he believed to be mainly maltose.
Emulsin
itself
does not hydrolyze maltose, and, according to

*
[6], 17, 225. t J. Chem. Soc., 73, 634; 83, 578. j Ber., 34, 600, 2206, 3810. Proc. Roy. Soc. (1905), 76 B, 592.
Arm-
J.
Pharm. Chim.
THE DISACCHARIDES
enzymic syntheses an isomeric sugar from the one which the enzyme itself hydrolyzes.
strong, in
Sugar.
705
is
obtained different
Thus:
Enzyme.
Product of
reaction.
1
1
maltose
isomaltose
2 d-glucose 2 d-glucose
is
+ +
maltoglucase
=
=
-j-
emulsin
-j-
=1
2 d-glucose 1 isomaltose 2 d-glucose maltose
Armstrong formed by the action of concentrated hydrochloric acid upon glucose (see under isomaltose). The products of this condensation after neutralization were treated with emulsin, which hydrolyzed the isomaltose, and then with Saccharomyces Marxianus, which fermented the glucose but not the maltose. The disaccharide remaining in solution gave an osazone corresponding to that of maltose; this and the biological behavior of
the sugar are strong indications of the formation of maltose. The question must be regarded, however, as unsettled until the sugar has
of the opinion that
both maltose and isomaltose are
been actually isolated in its pure crystalline form. The hydrolyzing enzymes undoubtedly exercise a synthetic action in the living cell, but the conditions under which this is accomplished are not understood sufficiently as yet to enable the chemist to control
the reaction in the laboratory.
ISOMALTOSE, C^HÔu.
No
other sugar has given rise to so
much
difference of opinion and uncertainty as isomaltose, a circumstance due to the fact that the so-called isomaltoses of different investi-
gators are in isomaltose was
all
probability
different
compounds.
The name
first
given
by Fischer*
to a synthetic disaccharide
prepared as follows.
One hundred grams of d-glucose were dissolved in Preparation. 400 gms. of cold fuming hydrochloric acid and the solution maintained for 15 hours at 10 to 15 C.; 4 kgs. of absolute alcohol were then added, the solution filtered from precipitated dextrins (reversion products) and the filtrate treated with a large excess of ether. The precipitate was filtered off, washed with alcohol and ether, pressed between filter paper, dissolved in a little water, the solution carefully neutralized with sodium carbonate, any alcohol and ether expelled by gentle warming and the exThe uncess of d-glucose removed by fermenting with yeast at 30 C. fermented residue (30 to 35 gms.) was dissolved in about 150 c.c. of water, exactly neutralized, and then heated with a solution of phenylhydrazine (30 gms.) in 50 per cent acetic acid (20 gms.) for 1| hours upon the water bath. The hot solution was then filtered from the slight
*
Ber., 23, 3687.
706
SUGAR ANALYSIS
upon cooling deposited
crystals
deposit of d-glucose-osazone; the filtrate

of isomaltose-osazone,
which differed from maltose-osazone by its greater and in water by its lower melting point, 150 C. solubility Theories Regarding the Formation of Isomaltose. Armstrong,
is
as previously mentioned, believes that maltose, as well as isomaltose, formed in the above synthesis. The maltose by Fischer's method of
is destroyed, however, by the action of the yeast. Isomaltose was also obtained by Scheibler and Mittelmeier* by the action of strong acids upon starch, the glucose which is first formed
purification
being afterwards recondensed to form isomaltose and other reversion products. The isomaltose thus formed is no doubt similar to that of
Fischer.
Isomaltose is also believed by Lintner,f Prior, J Albert and many other investigators to be formed during the diastatic conversion of starch. Opinions differ, however, as to whether this isomaltose is
formed before or after maltose; the following schemes illustrate a few of the numerous theories which have been proposed in this connection:
Starch Starch
Starch
>
amylodextrin
amylodextrin
isomaltose ~ isomaltose
> >
maltose. maltose.
> >
amylodextrin
||
* maltodextrin > maltose

.
.
maltose.
d-glucose
>
isomaltose.
Lintner and Dull prepared their isomaltose by saccharifying starch paste with malt extract at 70 C. The solution was then evaporated to a sirup, treated with strong alcohol, filtered from precipitated dextrin and the filtrate evaporated to expel alcohol; the d-glucose and maltose
were then fermented away with yeast, the solution clarified with bone black, evaporated to a sirup, treated again with strong alcohol to In this precipitate remaining dextrins and the filtrate evaporated. manner a white amorphous hygroscopic residue was obtained, which corresponded to the formula and molecular weight of C^HÔn + H 2 0. The substance was easily soluble in water and 80 per cent alcohol, and
showed
in
aqueous solution a
specific rotation of [a] D
= -+- 139
to
+ 140.
The
yield of isomaltose
by
this
method was about 20 per cent the
weight of starch.
Lintner and Dull believe that the hydrolysis of starch consists in a change of amylodextrin, or soluble starch, into lower dextrins which are then transformed into isomaltose and the
latter in turn into maltose.
*
Ber., 24, 301.
Chem.
II
Centralbl. (1894), 1131.
Chem.
Ztg., 16, 15.
Ber., 26, 2540.
t Z.
angew. Chem. (1892), 312, 872.
THE DISACCHARIDES
707
Ling and Baker,* repeating the work of Lintner and Dull, obtained a residue which gave with phenylhydrazine a mixture of osazones, cor-
and Baker
responding to d-glucose, maltose and an unknown trisaccharide. Ling also showed that a mixture of maltose and dextrin gave a
crystallizable osazone
which resembled
in
every
way
the so-called
Isomaltose-osazone.
and rnany other investigators Ost,| Ulrich,| Brown and Morris deny the formation of isomaltose during the diastatic conversion of starch and claim that the compound so designated is only a mixture
also of maltose
with different dextrins.
isomaltose prepared
by
his
Lintner claims, however, that the method, although not absolutely pure, is
,
sufficiently so to justify his conclusions as to its formation.
Emmerling and Armstrong regarding the formation of isomaltose by action of maltoglucase upon glucose have already been
of
The views
mentioned.
It
is
(See page 704.)
impossible to review in greater detail the copious literature upon isomaltose. No two authorities hold exactly the same opinion and the case is only an additional example of the lack of knowledge
which
still prevails regarding the different stages of starch conversion. Properties of Isomaltose. Isomaltose, as prepared by different
investigators using different methods, shows certain differences in physical and chemical properties. All preparations of the sugar reduce
Fehling's solution, Fischer's isomaltose having a reducing power 66 per cent and Lintner's 80 per cent of that of maltose. All preparations of the sugar upon heating with acids are hydrolyzed into d-glucose.
unfermented by yeast; that of Lintner in presis fermented but with considerable difficulty. The osazone test for isomaltose is regarded as the most characteristic, the greater solubility and lower melting point The distinguishing the osazone of isomaltose from that of maltose. melting point of Fischer's isomaltose-osazone on rapid heating is 158 C. The osazone of .Lintner's isomaltose Ost, however, gives 145 C. melts between 145 C. and 155 C. The osazone of both Fischer's and Lintner's isomaltose corresponds to the formula C24H32N40g. The fact that maltose in presence of impurities gives an osazone of
Fischer's isomaltose
is
ence of considerable yeast Tests for Isomaltose.
the same melting point greatly lessens the value of the osazone test for
isomaltose.
* t
I!
||
J.
Chem. Soc. Trans. (1895), 43, Chem. Ztg., 19, 1504; 20, 762.
fuller
702, 739.
For
" accounts of both maltose and isomaltose see Lippmann's Chemie
Chem. Ztg., 19, 1527. Chem. News, 72, 45.
der Zuckerarten" and Sykes and Ling's "Principles and Practice of Brewing.?
708
SUGAR ANALYSIS
Milk sugar.
Lactobiose.
LACTOSE.
HaOu
+ H 0.
2
Lactose is a sugar of distinctly animal origin, no Occurrence. well-authenticated evidence as yet existing of its occurrence in the vegetable kingdom. The sugar is formed in the lacteal glands of all
is built up from the glucose of the blood, although the which this synthesis takes place is not understood. Lactose is found in milk in amounts varying from less than 1 per cent to over 12 per cent, according to the kind of animal, period of lactation and other factors. The percentage of lactose in the milk of different animals
mammals and
in
manner
is
given in the following table:

Animal.
THE DISACCHARIDES
The
is
709
formed, albuminoids are coagulated and and other suspended impurities mechanically precipitated. The precipitate is filtered off and the filtrate saturated with carbon dioxide, filtered, evaporated, boiled to grain and centrifuged in the same manner
fat
of lime at 20 degrees Be. soluble calcium phosphate
free lactic acid is thus neutralized, in-
as for beet sugar manufacture. The yield of crystallized milk sugar this method is about 3.4 cent of the whey. by per
Properties of Lactose.
sists of large
Ci2H 2 20n
Milk sugar, as ordinarily prepared, conrhombic hemihedral crystals corresponding to the formula H2 0. The water of crystallization is given up with con-
siderable difficulty. The best method of dehydration is to precipitate a hot concentrated aqueous solution with 5 volumes of absolute
alcohol.
The
fine crystalline
powder thus obtained

,
is
dried
first
at
100 C., and then at 127 to 130 C when the last traces of water are given off without change in color or other evidences of decomposition.
Lactose hydrate is soluble in 5.87 parts of water at 10 G. and 2.5 parts of water at 100C.; it easily forms supersaturated solutions.
ether.
The sugar is insoluble The presence of
free alkali,
in absolute ethyl and methyl alcohols and in and of different salts of the alkalies,
increases the solubility of lactose in sucrose.
much
[a]
the same
manner
as with
Lactose hydrate
is
dextrorotatory,
= 36. The influence practically constant for concentrations up to c of temperature upon the [a] of lactose has already been referred to. D
The specific rotation of lactose anhydride is +55. 3.
is
=+
52.50 which value
is
(The same value
52.50 X ffrj.) [a] D of the hydrate, Freshly prepared solutions of lactose hydrate exhibit mutarotation. Tollens and Parcus* found for a solution of 4.841 gms. lactose hydrate dissolved to 100 c.c. the following values:
obtained by calculation from the
Time
after solution.
710
SUGAR ANALYSIS
Upon rapidly evaporating Low-Rotating Modification of Lactose. of 2 to 3 gms. of ordinary lactose hydrate in 10 c.c. of water solution a in a platinum dish and drying the residue at 98 C., Schmoger* obtained a form of lactose which showed after solution a rotation of
34.4 (calculated to Ci 2 22 Oii 2 O) but after 24 hours' standthe normal constant value of +52.5. to increased ing Erdmannf obtained the same form of lactose by rapidly boiling down a solution
only
+H
hydrate in a metal dish until the supersaturated solution a porous mass of small water-free crystals. Tansuddenly ret t obtained the low-rotating milk sugar (his so-called 7 modification)
of lactose
solidified to
by rapidly evaporating a solution of ordinary lactose (Tanret's so-called a modification) at 108 C., drying the crystals over concentrated sulphuric acid, dissolving rapidly in 3 parts of water and precipitating at
once with a large excess of alcohol. This process, after repeating several times, gives the pure 7 modification in the form of water-free crystals,
soluble in 2.2 parts of water at 15 C., which solution, however, on standing deposits crystals of the ordinary hydrate. Tanret's 7 milk
sugar 5 minutes after solution gave [a] D = 34.5, which value slowly increased upon standing to that of constant rotation. The change was
Tanret obtained an apparent third modification of lactose by allowing a concentrated solution of the hydrate to crystallize at exactly 85 to 86 C., or by treating the warm solution with 3 to 4 times its amount of absolute alcohol. Crystals were obtained, corresponding to the formula C ]2 H 22 On + J H 2 O, which gave immediately after solution in water the constant rotation +55.3
(calculated to the anhydride Ci 2
effected immediately Tanret's ^-lactose.
by adding a
trace of alkali.
22
On).
Hudson Upon
the Modifications of Lactose.

its
Hudson
in a recent
study of lactose and
modifications
came
to the conclusion that
Tanret's so-called or constant rotating lactose was not a definite compound but a mechanical mixture of the high and low rotating forms. The same conclusion was arrived at by Roux and also by Trey-TT Hudson, Roux, and Trey all confirm the earlier observation of Erd~ mann that the velocities of mutarotation for the high and low rotating forms of lactose are the same in value, and draw from this the conclusion that the two changes in rotation are only opposite parts of one
||
balanced reaction, which in its ultimate form may be expressed by the equation a-lactose <= /3-lactose. (The symbol (3 is given at present to
*
Ber., 13, 1915.

II
Z. physik.
t Ber., 13, 2180.
% Bull. soc. chim.
[3],
13, 625.
IF
Chem., 44, 487. Ann. chim. phys. [7], 30, 422 (1906). Z. physik. Chem., 46, 620.
THE DISACCHARIDES
the low rotating modifications of
Tanret.)
711
all sugars instead of 7 as first used by This identity in mutarotation for the high and low rotating
is
forms of lactose
shown by the following

TABLE CI
C. of Lactose Hydrate
results of
Hudson
*
:
Mutarotation at
and
fi-Lactose
Anhydride
Time
in hours.
712
SUGAR ANALYSIS
oxidized to lacto-
By action of bromine in aqueous solution lactose is

bionic acid.
The
latter
was
first
obtained by Fischer and Meyer,* as
a colorless sirup soluble in water, less soluble in alcohol but insoluble Lactobionic acid is hydrolyzed by dilute acids into d-galacin ether. This reaction and the reducing properties tose and d-gluconic acid. of lactose indicate the presence of a free aldehyde group attached to a
glucose residue. follows
:
The oxidation
of lactobionic acid
is
explained as
+O=
d-Galactose
radical
Cs
d-Galactose
radical
d-Glucose
radical
d-Gluconic acid
radical
Lactose
Lactobionic acid.
Lactobionic acid upon oxidation with hydrogen peroxide in presence of basic ferric acetate is degraded to galactoarabinose (p. 644).
Oxidation of lactose with nitric acid produces d-saccharic acid by

oxidation of the d-glucose radical, and mucic acid by oxidation of the The discovery of mucic acid as a result of this d-galactose radical.
oxidation was
lactose
made by
Scheele in 1780.
by the following method of
The yield of mucic acid from Kent and Tollensf is about 40 per
cent:
gr.;
100 gms. of pulverized lactose are dissolved in 1200 gms. nitric acid of 1.15 sp. the solution is evaporated to between 150 c.c. and 200 c.c. upon the water
filtered off after
bath and, after cooling, 200 c.c. of water added. 24 hours' standing.
The
crystals of
mucic acid are
Action of Alkalies. Lactose upon heating with dilute alkaliesj undergoes an almost complete loss of optical activity, the sugar molecule undergoing partial hydrolysis and rearrangement with formation of
d-galactose, pseudotagatose
sition.
and other products of unknown compoheating with concentrated alkalies lactose solutions are colored brown, the lactose being broken up into lower decomposition
Upon
among which lactic acid occurs in greatest amount. Lactose exposed to the action of alkalies in the sunlight yields according to Duclaux over 50 per cent lactic acid.
products
Lactose upon long treatment with calcium hydrate

into isosaccharinic acid.
is
converted
conducted as follows: A water is treated with 450 gms. calcium hydroxide and allowed to stand
*
reaction according to Kiliani is best cold solution of 1 kg. milk sugar in 9 liters of
The
Ber., 22, 361.
t Z. Ver.

14, 156, 203.
Lobry de Bruyn and van Ekenstein, Rec. Trav. Pays-Bas,

Ber., 16, 2625; 18, 631.
THE DISACCHARIDES
at
clear
713
room temperature with frequent shaking for 5 to 6 weeks. The brown solution is saturated with carbon dioxide, filtered from calcium carbonate, heated to boiling, again filtered and concentrated to 2 liters. Calcium isosaccharinate crystallizes out and calcium metasaccharinate remains in solution. The precipitate of the former is filtered off, washed with cold water and dried. A weighed quantity of the salt is then dissolved in water and decomposed with an exact The filtered solution contains free isosacamount of oxalic acid. charinic acid, C 6 Hi 2 O 6 which upon evaporation splits off water, and
,
crystallizes out as its lactone or isosaccharin,
Hi
5.
Isosaccharin consists of beautiful double-refracting crystals, easily It melts at 95 C. and sublimes soluble in water, alcohol and ether.
It is dextrorotatory ([oj^ = without decomposition. 63), fermented by yeast and does not reduce Fehling's solution.
is
not
Kiliani's isosaccharin
is
identical with
Nef's a-isosaccharin, ob(p. 603),
tained
by the action
of
sodium hydroxide upon d-galactose

:
and
has the following structural formula
CH OH
2
HCH HOH C -COH

2
Hydrolysis of Lactose by Acids.
Lactose upon heating on a

is
boiling water bath with 4 parts of 2 per cent sulphuric acid * lyzed into equal parts of d-glucose and d-galactose
:
hydro-
C H On
12
22
+H
= C
12
CH
6
12
6.
Lactose
d-Glucose
d-Galactose.
The hydrolysis may also be carried out with hydrochloric acid, the reaction proceeding at ordinary temperature with sufficient concentration of acid. Urechf found that a solution containing 12 gms. lactose
and 32 gms. hydrochloric acid to 100 c.c. was hydrolyzed almost completely after 12 hours' standing at 23 C. The hydrolysis of lactose by means of acids proceeds with much greater difficulty than that As in the case of maltose the prolonged action of sucrose or maltose. of acid causes a slight loss of reducing sugar due to formation of levulinic acid,
substances, reversion products, etc. In so far as the different yeasts, Fermentation of Lactose.
humus
bacteria, moulds, etc., contain the lactose-splitting enzyme lactase, they ferment lactose in the same manner as d-glucose and d-galactose.
*
Ost, Ber., 23, 3006.
t Ber., 18, 3048.
714
SUGAR ANALYSIS
organisms which ferment d-glucose and d-galactose are without upon lactose, and in such cases it is generally supposed that
is
Many
action
lactase
absent.
But whether
in all lactose fermentations the sugar

is
must
first
be hydrolyzed by a lactase
uncertain, although
many
au-
thorities at present incline to this opinion.
Ordinary baker's and brewer's yeasts exercise no pronounced fermentation upon lactose. Hansen,* in fact, found that none of the common alcohol-producing yeasts in pure culture was able to ferment
Invertase, maltase, zymase and the other enzymes of ordinary yeast have in fact no action upon lactose. The action of different yeasts upon lactose and other sugars is given by Fischerf and
lactose.
Thierfelder in Table
GIL
TABLE CII
Showing Action of Yeasts Upon Different Sugars
Kind
of yeast.
THE DISACCHARIDES
Lactase.
715
best prepared according to Fischer* by washing (the granular concretions of yeasts and bacteria which constitute the Kefir ferment) with water, grinding the air-dried mass
is
Lactase
Kefir corns
with pulverized glass and extracting with water. Lactase is much less sensitive to the paralyzing action of alcohol, acids, etc., than invertase, amylase, maltase and other enzymes.
lactase
its occurrence in the Torulaceae and other organisms found in several species of moulds and fungi and also in higher plants. The crude emulsin, obtained by extracting almonds and the seeds of other Rosaceae, contains a lactase, although the emulsin from Aspergillus niger, the cherry laurel and several other sources has no hydrolyzing action upon lactose. Lactases are very abundant in the animal kingdom and this would be expected from the importance which lactose plays in animal nutrition, Lactases have been found in the especially during the nursing period. mucous membranes of the stomach and intestines of newly born infants and in the same tissues of the young of most mammals. Lactase
In addition to
is
also occurs in the
membranes
of the digestive tract of older animals
and
is
found in preparations of such enzymes as ptyalin, pepsin, trypsin,

It is a significant fact that the secretion of lactase is in-
rennet, etc.
tensified
increasing the amount of milk or milk sugar in the food. and Butyric Fermentations. Lactose is more easily subject In fact, to the lactic and butyric fermentations than any other sugar. which lactic from the acid all produce nearly organisms d-glucose and
by
Lactic
sucrose ferment lactose in the
same way. On the other hand, there are a very large number of organisms which produce lactic acid from lactose but not from sucrose or glucose.
The
lactic
fermentation to secure best results must be conducted in
presence of suitable nutrients and also in presence of calcium carbonate (or similar substance) to neutralize the free acid as soon as formed.
of much free acid checks the fermentation, so that in milk, for example, the percentage of free lactic acid under ordinary conIn presence of sufficient ditions never exceeds 0.5 to 0.6 per cent.
The presence
calcium carbonate the fermentation of lactose into lactic acid under

favorable conditions
may
be almost quantitative.
organisms differ greatly in their action upon lactose.. While the ordinary optimum temperature is about 30 C., some bacteria produce fermentation as low as 10 C., and others as high as 52 C. Some lactic organisms are retarded and others stimulactic
The various
lated
by the influence
of atmospheric oxygen.
*
The
lactic bacteria also
Ber., 27, 3479.
716
differ as to
SUGAR ANALYSIS
the kind of lactic acid which they produce, some organisms form d-lactic acid, others 1-lactic acid, others d, 1-lactic acid and others mixtures of d- and 1-lactic acids in varying amounts.
The
and
literature
upon the
lactic acid bacteria is exceedingly copious
for a review of the subject reference should
be made to the works
of Lafar,* J6rgensen,f
Emmerling,J and
others.
In the butyric fermentation of lactose the butyric acid may be formed either primarily from the lactose or secondarily from the lactic A large class of both aerobic acid produced by the lactic fermentation. and anaerobic bacteria ferment lactose with production of butyric acid, but the action of only a few of these has been studied. In addition to
butyric acid, formic, acetic, propionic, valeric and succinic acids, butyl
alcohol, acetone, carbon dioxide, hydrogen and methane have found among the products of different butyric fermentations.
been
large
viscous gum.
bacteria of this class have been especially studied in connection with the slimy or ropy fermentation of milk. The Leu-
number The
of
organisms ferment lactose with formation of a
and many other organisms which form dextran from sucrose do not exercise this action upon lactose.
conostoc mesenterioides
Lactose, the same as maltose, contains Compounds of Lactose. a reactive aldehyde group and forms hydrazones, osazones, methyl lactosides, ureides, mercaptals, etc., in the same manner as other re-
ducing sugars.
The most important

is
of these
compounds from the

:
6 5)2 lactose-phenylosazone, C^HôOg ( which was first prepared by Fischer by heating 1 part lactose, 1J parts phenylhydrazine chloride, 2 parts sodium acetate and 30 parts water
analytical standpoint
N NHC H
The osazone separates from the cold solution in the form of yellow needles, which after recrystallizing melt at 200 C. Lactose phenylosazone is soluble in 80 to 90 parts boiling water, in hot alcohol and in hot glacial acetic acid; it is insoluble in ether, benzol and chloroform. Lactose forms with nitric acid, in presence of ice-cold strong sulphuric acid, tri-, tetra-, penta,- hexa- and octonitrates; and with acetic
for 1J hours.
anhydride mono-,
is
di-,
tetra-,
hexa- and octacetates.

||
The
octacetate
acetate, and is obtained by heating 5 gms. lactose with 20 gms. acetic anhydride and 5 gms. water-free sodium
the best
*
t
known
Technische Mykologie." Microorganismen der Garungsindustrie." " t Zersetzung Stickstoffreier organ. Substanzen durch Bacterien," Braunschweig (1902), pp. 25-84.
"
Ber., 17, 579; 20, 821.
II
"
Schmoger, Ber., 25, 1452.
THE DISACCHARIDES
acetate just to boiling and then pouring the mass into water.
717
The prefrom alcohol. Lactose octacetate recrystallized cipitated consists of white crystals of the formula Ci 2 Hi4(C2H 3 O) 8 Oii, and meltcompound
is
The ing according to different observers between 86 C. and 106 C. variation in melting point is attributed by some chemists to the existence of
pound
sates,
two isomers. The rotation of the carefully purified com=' 3.5 (in chloroform). given by Schmoger as [a] D Lactose forms with alkalies and alkaline earths a number of lactois
none
of which, however, has
any analytical or technical imporacid lactose forms a nitrile, which
tance.
Upon treatment with hydrocyanic
yields after saponification lactose carboxylic acid; the latter consists of
a colorless sirup and gives upon hydrolysis d-galactose and a-glucoheptonic acid.
C H 05CH-0-C H
5 10
12
COOH + H
= CH
6
12
+C H
6
13
COOH.
Lactose carboxylic acid
d-Galactose
a-Glucoheptonic acid.
Characteristic qualitative tests for lactose in are of other wanting. The formation of mucic acid upon sugars presence oxidation with nitric acid is a valuable confirmatory test; although it
Tests for Lactose.
must always be borne in mind that the same reaction is also given by d- and 1-galactose, galactonic acid and galactan. Separation of lactose osazone, after careful recrystallization and purification, offers a In case several reducing sugars fairly reliable means of identification.
are present, the mixture of osazones should be heated with boiling
water and
place
filtered;
saccharides will be found in the
the osazones of lactose, maltose and other difiltrate, from which crystallization takes
upon
cooling.
In case of mixtures, the destruction of d-glucose, d-fructose, d-mannose, sucrose, maltose and other fermentable sugars by means of pure cultures of yeasts which do not ferment lactose (Table CII) may be
employed to advantage before making

Configuration.
lished
tests for lactose.
The configuration of lactose has not been estabwith certainty. The following constitutional formula proposed
to
by Fischer* answers, however,

reactions of lactose
:
most
of the chemical properties
and
H H H OHH OHH HOH C-C-C-i-C-C-0-C-C-C-C-C-CHO H OHOHH OH inH H OHH
d-Galactose radical
d-Glucose radical
* "
Untersuchungen uber Kohlenhydrate
"
(1909), p. 92.
718
Synthesis.
SUGAR ANALYSIS
The synthesis of lactose has not yet been accomplished by means of enzymes.
either chemically or
Isolactose.
C^HÔu.
The name was given by
Isolactose has not been found in nature.
Fischer and Armstrong* to a disaccharide, which they obtained through the synthetic action of Kefir lactase upon a concentrated solution of
equal parts d-glucose and d-galactose. Fifty grams of finely shredded Kefir corns were shaken up with 300 c.c. water and 5 c.c. toluol for 48 hours at ordinary temperature;
200
10
c.c.
c.c.
of the extract, 100 gms. each of d-glucose and d-galactose, and of toluol were placed in a closed flask and allowed to stand 15
days at 35 C.
After diluting with
and
cooling, the filtered solution
1 volume of water, boiling 10 minutes was fermented with top yeast to re-
move
the residual d-glucose and d-galactose.
The
isolactose remain-
ing in solution
was separated only

Trehabiose.
in
form of isolactose-phenylosazone,
melting at 190 to 193 C.
C24 H32N 4 O9,
which consisted of
fine needles
TREHALOSE.
Occurrence.
Mycose.
Mushroom
2
sugar.
Ci 2
22
On
+ 2 H 0.
is a disaccharide discovered by Wiggersf growth produced by the fungus Claviceps purpurea upon rye and other grasses), and later found to occur in nearly Muntz { found in the dry substance of all fungi and mushrooms. Agaricus muscarius, A. sulfureus, A. sambucinus, Fungus sambuci and
Trehalose
in ergot (the sclerotium
Boletus cyanescens as much as 10 per cent trehalose. Bourquelot also found trehalose in varying amounts in Sclerotinia tuberosa, Phallus impudicus, Boletus satanas and in 36 other varieties of Boletus,
Amanita, Pholiota, Hypholoma and Lactarius. Trehalose is unequally distributed through the several tissues of mushrooms; the stems of Boletus edulis, for example, were found by Bourquelot to contain 2.45 per cent
The quantrehalose, the caps 1.38 per cent and the pores none at all. tity of trehalose also varies according to the period of vegetation, being After the mushroom is greatest just before the formation of spores.
picked, the trehalose
is rapidly hydrolyzed by the enzyme trehalase into the latter d-glucose, being afterwards reduced through some unknown to mannite. biological process Trehala-manna and Trehalum. Trehalose was also found by
Berthelot
*
||
in the so-called
Trehala-manna, a drug long used in the

Compt. rend., 108, 568; 111, 578; Ann. chim. phys. [3], 56, 272.
117, 826.
Ber., 35, 3144.

||
t Ann., 1, 173 (1832).

J
Compt.
rend., 79, 1182.
THE DISACCHARIDES
Orient,
719
and consisting of the cocoon or gall-like concretions formed by a species of beetle upon certain spiny plants of Syria and Persia. Trehala-manna also contains a polysaccharide trehalum, discovered by Guibourt* and later investigated by Scheibler and Mittelmeier.f Trehala-manna is first extracted with successive quantities of boiling alcohol to remove the trehalose and then with boiling water. The hot-water extract is filtered through a hot-water funnel and the trehalum separates from the cold filtrate as a white tasteless crystalline
powder, with a composition corresponding to the provisional formula C24H 42 02i. Trehalum is soluble in 56 parts of boiling water, is dextro179), and is hydrolyzed by hydrochloric or sulphuric rotatory ([a] I) = acids into d-glucose; it is not reducing and forms no compound with phenylhydrazine. Solid trehalum is colored by alcoholic iodine solution
violet
and solutions
of
trehalum wine red.

it
Trehalum has a
certain
possibly bears the same relation to trehalose that starch bears to maltose.
resemblance to starch
and
Preparation of Trehalose.
Finely shredded trehala-manna or
mushrooms (freshly picked) are boiled with strong alcohol and the filtered extract set aside to cool. Crystals of trehalose will usually
separate immediately; if crystallization does not take place the alcoholic extract is concentrated by evaporation and again set aside.
Trehalose, as ordinarily prepared, is Obtained in Properties. the form of large colorless rhombic crystals having the formula
The crystals have a sweet taste, are soluble in 2 water and in 100 parts of hot alcohol, but are insoluble in The hydrate begins to melt in its water of crystallization at ether. 94 C., and liquefaction is complete between 96.5 C. and 97.5 C. The water of crystallization is lost at about 130 C. and the trehalose anhydride thus obtained melts at about 200 C. Trehalose is strongly dextrorotatory; SchukowJ found for the hyCi 2
2
H 2Oii + 2 H O.
1.7 parts of
drate
(c
7.282) [a]%
=; -f-
178.3,
the anhydride would be [a]g
from which the calculated value for =+197.1. Apping determined for the
Mutarotation is not observed. 197.28. anhydride [a] D = Reactions. Trehalose, like sucrose, does not reduce Fehling's solution. The absence of aldehyde or ketone properties is also shown by the failure of trehalose to form hydrazones or osazones, by its resistance towards solutions of boiling alkalies, and by the non-forma-
tion of acid oxidation products in aqueous solution by means of bromine at ordinary temperature. Upon warming with dilute nitric acid
*
Compt.
rend., 46, 1213.
t Z. Ver.
Deut. Zuckerind, 60, 818.
t Ber., 26, 1331.
Dissertation, Dorpat., 1885.
720
trehalose
is
SUGAR ANALYSIS
oxidized to d-saccharic acid
and by strong
nitric acid to
oxalic acid.
Upon
hours, trehalose
heating with dilute hydrochloric or sulphuric acids for several is hydrolyzed into 2 molecules of d-glucose.
v-a2-H22v)ii -j- -H^jO Trehalose
2
d-Glucose.
accomplished only with considerable difficulty. hydrolysis 6 hours' boiling with 5 per cent sulphuric Winterstein* to According acid is necessary to secure complete conversion; the yield of d-glucose
is
The
obtained at the end of this time was 99.45 per cent of the theoretical. Trehalose forms a number of acetates upon heating with acetic
anhydride and a number of benzoates upon treatment with benzoyl chloride. Compounds of trehalose with calcium, strontium and lead
is not readily fermented by ordinary The sugar is fermented, however, by a numbaker's or brewer's yeast. ber of wild yeasts; according to Dubourgf a large number of yeasts
have also been prepared. Fermentation. Trehalose
can ferment trehalose after special methods of cultivation and adaptaA number of moulds, as Mucor mucedo, ferment trehalose in absence of air with production of alcohol. The fermentation of trehalose is apparently conditioned by the presence of a special hydrolyzing enzyme trehalase, which was first isolated by Bourquelot t from a culture of Aspergillus niger at the time of spore formation. The mould was distintegrated by grinding with
tion.
in a
sand, dehydrated by washing with 95 per cent alcohol and then dried vacuum at low temperature. The dried mass was then extracted
with water, and the trehalase precipitated from the filtered extract by strong alcohol. The purified enzyme consisted of a white amorphous
substance, easily soluble in water and having below 53
C. a strong hydrolytic action. Its activity was destroyed by heating to 63 C. Trehalase is found, according to Bourquelot, in other moulds and in
It has also been dehigher fungi, such as Morchella and Polyporus. tected in green malt. In the animal kingdom its presence has been reported in human urine, in the pancreas and intestines of rabbits, in the
intestines of calves, horses, etc.,

fishes.
and
in the blood
serum
of certain
Emulsin, invertase, diastase and ptyalin have no hydrolytic action
upon
trehalose.
Tests.
*
No
reliable qualitative tests are
known
J
for the detection

if
of trehalose in mixture with other sugars.

Ber., 26, 3094.
f
According to Bourquelot,
Compt.
rend., 117, 826.
Compt.
rend., 128, 440.
THE DISACCHARIDES
a
glass plate
721
be gently rubbed with a crystal of trehalose and a drop of sirup containing trehalose be spread over the rubbed area, crystallization will develop and the lines of contact, where the trehalose crystal touched the glass, will appear visible.
Configuration.
to the absence of reducing properties, it is that the supposed aldehyde groups of the two glucose radicals are combined together. The following arrangement has been proposed:
been established.
The Owing
constitutional formula of trehalose has not
H H OHH H HOH C-C-C-C-C-C\ OH H OH

2
I
I
\
>o
H H OHH H/ HOH C-C-C-C-C-C H OH OH

2
I
|
MELIBIOSE.
Occurrence.
Ci 2
H 22 O n 2 H2 0. This disaccharide has not been found in nature as
yet in the free condition. It was first prepared by Scheibler and Mittelmeier* by a partial hydrolysis of the trisaccharide raffinose by means of mineral acids (see page 737)
.
CisH32 Oi6
Raffinose
+ H O = d-Fructose CeHÔe + Ci H 2On.

2
Melibiose.
The same hydrolysis is effected by means of invertase. pure culture of top-fermentation yeast will hydrolyze raffinose into melibiose and d-fructose and, after fermenting the latter, leave a solution containing melibiose. Bottom-fermentation yeast cannot be used owing to the occurrence of an enzyme, melibiase, which hydrolyzes melibiose as fast
as
it is
formed.
For the preparation of melibiose either of the methods proposed by Bauf may be used. A steriPreparation of Melibiose from Raffinose by Fermentation. of c.c. lized solution, containing 20 gms. raffinose in 250 water, is
Preparation.
fermented with 30 gms. of a pressed pure culture of Frohberg top-fermentation yeast at 31 C. for one day. The solution is filtered, sterilized and again fermented with 10 gms. of yeast at 31 C. for several days. The filtered solution, after decolorizing by means of bone black, is
evaporated, the hot sirup poured into hot 95 per cent alcohol, and the cold solution, after decanting from precipitated impurities, treated with
an excess of ether.
*
The impure
melibiose,
t
which
Chem.
is
thus precipitated,
Ber., 22, 1678; 23, 1438.
Ztg., 26, 69.
722
is
SUGAR ANALYSIS
C. as its dissolved in 70 per cent alcohol and precipitated again at barium compound by adding cold aqueous barium hydroxide solution and cold* 92 per cent alcohol. The barium melibiate is filtered off, washed with cold alcohol, pressed, suspended in water and decomposed by means of carbon dioxide. Any barium remaining in solution is precipitated by adding the exact amount of dilute- sulphuric acid (avoiding even the slightest excess) and the filtrate evaporated below 80 C. to a sirup; 92 per cent alcohol is added until the alcoholic strength of the diluted sirup is 70 per cent and ether is added just beyond the point of
precipitation.
The ether-alcohol
solution after filtration
is
set aside
and
gradually deposits crystals of melibiose, which recrystallizing from 78 per cent alcohol.
may
be purified by
Preparation of Melibiose from Raffinose by Hydrolysis with Acid. 10 to 20 per cent solution of raffinose is boiled with 2 per cent acetic
the solution
is
concentrated in a porcelain dish to a thick sirup rubbed up with 2 volumes of 95 per cent alcohol. which is decanted and ether added with shaking to the solution The alcoholic
acid;
after cooling is
point of permanent turbidity; the solution after standing 2 days is filtered from the precipitate and allowed to stand for several weeks in a closed
Crystallization gradually takes place; the process may be hastened by priming with a little melibiose from a previous preparation. The crude melibiose thus obtained is purified by recrystallizing from
flask.
alcohol.
Melibiose is obtained from aqueous solution in the Properties. form of monoclinic crystals having the formula Ci 2 H 22 Oii + 2 H 2 0. The sugar has a mild sweet taste, and when warmed begins to melt in to 80 C. its water of crystallization at 75 Upon heating in a thin to C. all water is The anhydride is ex110 expelled. layer gradually and reabsorbs water from the air with great ceedingly hygroscopic
One gram of crystallized melibiose is dissolved by 0.4186 avidity. water at 17.5C., by 6.8137 gms. of methyl alcohol and by 175.67 of gms.
gms. of ethyl alcohol. Melibiose is strongly dextrorotatory. Bau found for the hydrate = 129.50, from which the value for the anhydride = 143.
MS
The sugar
exhibits mutarotation, the initial value, for [a] D being less than the constant reading ([a] after 5 minutes = 108.68).
Melibiose reduces Fehling's solution, the reducing power as C^HÔn being about 95 per cent that of maltose. Reduction with sodium amalgam gives a complex alcohol, melibiotite, Ci 2 24 On,
Reactions.
which consists of an easily soluble non-reducing sirup, and is hydrolyzed by acids into d-galactose and d-mannite. Upon heating with hydrochloric
THE DISACCHARIDES
or sulphuric acid melibiose
d-galactose.
is
723
slowly hydrolyzed into d-glucose and
CuHaOu
Melibiose
+H
= CH
6
12
+CH
6
12
6.
d-Glucose
d-Galactose.
Melibiose
lactose.
acids.
also,
is thus seen to yield the same products of hydrolysis as Hydrolysis is not effected by acetic, tartaric, citric or lactic Melibiose is rapidly hydrolyzed by the enzyme melibiase, and
but more slowly, by emulsin. Owing to the presence of a reactive aldehyde group melibiose forms a considerable number of hydrazones and osazones. The sugar also forms an octacetate upon boiling with acetic anhydride and sodium
consists of needle-shaped crystals with very having the formula CwHuCCkHaOÔii, only slightly soluble in water, soluble in alcohol, chloroform and benzol and showing dextrobitter taste,
acetate.
The compound
rotation
is readily fermented into alcohol and carbon dioxide by all bottom-fermentation yeasts, but not by pure cultures of the ordinary top yeasts. Certain exceptions to the latter rule have been noted in a few cases, but fermentation in these instances may have been due to contamination or to the unexplained phenomenon of transition by which a bottom yeast acquires top-fermentation characteristics, or vice versa. Pure cultures of Saaz and Frohberg top yeast, Saccharomyces ellipsoideus II, as well as other varieties, were found by Bau* to have no action upon melibiose even after 1 to 1 \ years; this property has been proposed by Bau as a means of distinguishing between top and bottom yeasts; the dangers of contamination and transition nullify somewhat the value of this test. A large number of wild yeasts, mycoderms, moulds and other
= 94.2.) ([a] D Fermentation. Melibiose
fungi ferment melibiose; in all such cases the presence of a special enzyme melibiase is assumed. Melibiase, as prepared by Bau from
bottom
tion;
yeast, can be heated to 110 C. (after drying) without destrucThe in solution, however, its activity is destroyed at 70 C.
for its action
is
optimum temperature
Tests.
about 50 C.
Reliable qualitative tests for melibiose in presence of other sugars are lacking. The best method of procedure in case of mixtures is to remove fructose, glucose and other sugars so far as possible
The melibiose may then be precipitated culture of top yeast. as its phenylosazone; the latter after recrystallization from hot water to 179 C. Melibioseconsists of fine yellow needles melting at 178
by a pure
*
Chem.
Ztg., 19, 1874; 21, 185.
724
SUGAR ANALYSIS
phenylosazone is decomposed by benzaldehyde into melibiosone, which is hydrolyzed by emulsin into d-galactose and d-glucosone. Oxidation of melibiose or its osone with nitric acid yields mucic acid, in
the
same manner
Synthesis.
as with lactose.
allowing acetochloro-d-galactose to react with d-glucose in alcoholic solution in presence of sodium alcoholate Fischer and Armstrong * obtained a disaccharide which reduced Fehling's
By
solution,
formed osazones similar to those of melibiose and gave other
The sugar could not be separated in reactions resembling this sugar. the crystalline form, but from the agreement in chemical reactions
and
in behavior towards yeasts
and enzymes
it
is
probably identical
with melibiose.
The above synthesis is of importance as it is apparently the first to be accomplished by purely chemical means for a natural disaccharide; the method is the same as that employed for other synthetic disaccharides,
and may be represented as
follows:
Cl
THE DISACCHARIDES
Turanose.
725
C^HÔn.
This disaccharide has not been found free in nature. The sugar was obtained by Alekhine * by a carefully controlled hydrolysis of the
trisaccharide melezitose (p. 740).
CisHÔie Melezitose
HÔ = CeHÔe
d-Glucose
C^HÔn.
Turanose.
About 5.5 gms. of anhydrous meleziPreparation and Properties. tose are carefully warmed with 100 c.c. of 1 per cent sulphuric acid upon the water bath until the specific rotation of the sugar has fallen to
M0
= ~^~ 63*
The
solution
is
then neutralized with barium carbonate,

After
purified
and the
nitrate treated with hot alcohol until turbidity develops.

is
cooling the precipitated sugar

lute alcohol.
by extracting with
boiling abso-
Turanose
is
obtained by the above method as a white amorphous
mass, having the formula C^HÔn; it is easily soluble in water and 65 to +68 methyl alcohol but not in absolute alcohol, [oj^ =
not fermentable to any great extent and reduces Fehling's solution about one-half as strong as d-glucose. Turanose upon prolonged heating with mineral acids is hydrolyzed
(c
30 to 35).
It is
according to Alekhine into 2 molecules of d-glucose. Tanret f has recently prepared turanose by hydrolyzing melezitose
with 20 per cent acetic acid for 2 hours on the water bath. The acetic acid was removed by shaking out with ether and the d-glucose by fermenting with yeast. The solution was evaporated to a sirup, purified
from glycerol and fatty acid fermentation products by shaking out with ether, and then extracted with boiling absolute alcohol. The filtrate on concentration and cooling yielded crystals of turanose contained one-half molecule alcohol of crystallization and melted at 60 to 66 C. Upon drying at 100 C. anhydrous turanose was (which obtained of the formula Ci 2 H 22 Oii and [a] D = + 71.8. The reducing power was 60 per cent that of d-glucose. According to Tanret turanose is hydrolyzed into one molecule each of d-glucose and d-fructose, turanose thus being a true isomer of sucrose. This observation cannot be reconciled with the rotation +51, obtained
by Alekhine
after hydrolyzing melezitose; additional investigations are
needed before a final decision can be reached. Turanose upon heating with a solution of phenylhydrazine forms a phenylosazone of the formula C^H^NA, consisting of fine yellow needles, melting at 215 to 220 C. upon rapid heating and soluble in
*
Ber., 22, 759;
Ann. chim. phys.

[3],
[6],
18, 532.
t Bull. soc.
chim.
36, 816.
726
SUGAR ANALYSIS
of fine crystals.
5 parts of hot water, which solution upon cooling sets to a jelly-like
mass
Gentiobiose.
C^HÔu.
In the com-
This disaccharide has not been found free in nature.

bined form
it
it has been obtained by Bourquelot and Herissey* by partial hydrolysis using invertase or very dilute sulphuric acid.
exists in the trisaccharide gentianose
from which
Gentianose
d-Fructose
Gentiobiose.
Ten grams of gentianose are warmed Preparation and Properties. upon the water bath with 100 c.c. of 0.2 per cent sulphuric acid for 30
minutes.
filtered
The
solution
is
and evaporated
in
vacuum
cooled, neutralized with calcium carbonate, to dryness. The residue is worked
up with absolute alcohol and then with 95 per cent alcohol until all fructose is removed; the crude gentiobiose is purified by crystallization. Gentiobiose, when crystallized from hot alcohol, is obtained in
water-free crystals which, after drying at 115 C., melt at 190 to 195 C. The sugar is dextrorotatory and shows the phenomenon of
less-rotation,
[a]^
=+9.61
(after solution).
The sugar
is
not
fer-
mented by top yeast and this property can be utilized for separation of While gentiogentiobiose from the hydrolytic products of gentianose.
is not hydrolyzed by invertase, it 2 molecules of d-glucose. Hydrolysis 3 per cent sulphuric acid.
biose
is is
easily split
also effected
up by emulsin into by heating with
Ci2H220n
Gentiobiose
-|-
HÔ =
2 CeHÔe.
d-Glucose.
Gentiobiose reduces Fehling's solution to about the same degree as maltose. Upon heating with phenylhydrazine an osazone is formed of
These reactions show that gentiobiose contains an aldehyde group in the free reactive condition.
melting point 142 C.
Cellose.
Cellobiose.
CuEÔu.
exist, so far as
This disaccharide does not

It is
known,
free in nature.
apparently formed, with other intermediary carbohydrates, in the
hydrolysis of cellulose to glucose by means of sulphuric acid, cellose thus bearing the same relation to cellulose as maltose bears to starch.
For the preparation f of cellose it is best to start Preparation. from cellose-octacetate, which is obtained by treatment of cellulose with acetic anhydride: 7.5 gms. of finely cut filter paper are thoroughly shaken in a 200-c.c. flask with 20 c.c. of acetic anhydride; after cooling
*
Compt.
rend., 132, 571; 136, 290, 399.
Skraup, Ber., 32, 2413.
THE DISACCHARIDES
to 70
727
C. the mass
is
c.c.
acetic anhydride
treated under constant shaking with a mixture of and 4 c.c. concentrated sulphuric acid warmed
to 70 C.,
of water.
alcohol. The cellose-octacetate thus prepared conwhite needles, melting at 228 C., and having a composition and molecular weight corresponding to the formula Ci2 Hi4(C 2 H 3 0) 8 Oii. The
sists of
on linen, from 95 per cent
and the yellowish brown solution poured into 500 to 700 c.c. The amorphous yellowish colored precipitate is filtered off washed with water, pressed and recrystallized several times
compound is soluble in hot 95 per cent alcohol, but insoluble in hot absolute alcohol, chloroform or benzol. The yield of cellose-octacetate from cellulose by this method is 26 to 27 per cent.
To prepare
cellose the finely pulverized octacetate is
moistened
with alcohol and then treated with successive portions of a 15 per cent solution of potassium hydroxide in strong alcohol, using 5 c.c. of alcoholic potassium hydroxide for each gram of cellose-octacetate. By this
treatment the octacetate
is
saponified
and the
Ci 2
22
cellose liberated.
3
12
H 4(C2H 0)
1
11
+ 8KOH =
Cellose-octacetate
Cellose
H O n + 8CH COOK. Potassium
acetate.
After 2 hours' standing the crude cellose, in the form of a granular powder, is filtered off, washed with absolute alcohol, dissolved in a little water and any free potassium hydroxide exactly neutralized with acetic The solution is then filtered, evaporated to a thin sirup, 1 part acid. absolute alcohol added and then ether to the point of turbidity. After
standing several hours in the cold the precipitate is filtered off, dissolved in a little hot water and then absolute alcohol added to the appearance The solution is then set aside in the cold when the celof turbidity.
lose will separate after long standing in the
crystals.
form of small microscopic

consists
Properties.
Cellose as prepared
by the above method
of a fine white crystalline
compound, which,
after drying at 100
C. in
a vacuum, has a composition agreeing with the formula Ci 2 H220n. The sugar melts at 225 C. under decomposition. It has a mild sweet
taste, is soluble in
8 parts of cold and 1.5 parts of hot water, but inCellose is dextrorotatory and shows soluble in alcohol and ether. 26.1 (10 minutes after soluthe phenomenon of less-rotation, [] = Upon heating on the water bath with tion) and +33.7 (constant).
5 per cent sulphuric acid for 7 hours cellose

cules of d-glucose.
is
hydrolyzed into 2 mole12
Ci 2
22
On
+HO
2
H O
6.
Cellose
d-Glucose.
Hydrolysis
is
also effected
by means
of emulsin.
728
Cellose
is
SUGAR ANALYSIS
not fermented by means of yeast.
The sugar reduces
Upon heating Fehling's solution somewhat stronger than maltose. with 2 parts water, 3 parts phenylhydrazine and 2 parts glacial acetic acid for 1 hour in a water bath and then adding water, cellose forms an
osazone which crystallizes in the form of yellow needles melting at 198 C. These reactions show that cellose contains an aldehyde group
in the free reactive condition.
Glucosido-galactose. This synthetic disaccharide was prepared by Fischer and Arm-
strong* from acetochloro-d-glucose and d-galactose (in alcoholic solution with sodium) following the same method described under the synthesis of melibiose.
by bottom
It
The sugar was obtained only in the sirupy form; it was fermented yeast, but not by top yeast, and was hydrolyzed by emulsin.
reduced Fehling's solution.
Phenylhydrazine gave an osazone C24H 32 4 09 consisting of yellow needles, melting at 172 to 174 C., and soluble in 120 parts of hot
water.
Galactosido-galactose. This synthetic disaccharide was prepared by Fischer and
Arm-
strong* from acetochloro-d-galactose and d-galactose (in alcoholic solution with sodium) following their general method as described under
melibiose.
The sugar was obtained only in the sirupy condition; it was fermented neither by top nor bottom yeast but was hydrolyzed by emulsin;
it
reduced Fehling's solution.
Phenylhydrazine gave an osazone C24H 32 4 09 consisting of yellow needles, melting at 173 to 175 C., and soluble in 110 parts of boiling
water.
Isotrehalose.
C^HÔu.
was prepared synthetically by Fischer and somewhat different process than that of Fischer and
This
disaccharide
Delbriickf by a
Armstrong. /3-Acetobromoglucose is allowed to react in dry ethereal solution with silver carbonate, traces of water being added at intervals to
promote the reaction

are united
by the elimination
In this manner two molecules of acetoglucose of bromine to form the octacetate of a

*
Ber., 36, 3144.
t Ber., 42, 2776.
THE DISACCHARIDES
disaccharide.
synthesis:
729
The
following equation illustrates the principle of the
,-CHBr
01
I
CHOAc
CHOAc
3
01
I
CHOAc
01
CHOAc CHOAc
*U H
CHOAc
+A&C0 =[_ iH
CHOAc
+ 2AgBr+ C0
CHOAc
CHOAc
CH OAc
2
CH OAc
2
CH OAc
2
Acetobromohexose
Disaccharide octacetate.
The
octacetate upon treatment with cold barium hydroxide solu-
tion yields barium acetate and the free disaccharide. Isotrehalose as obtained by this process consists of a white amor-
phous substance, without action upon Fehling's solution and levoThe absence of a free aldehyde group is a 39.4). rotatory ([a] D = necessary consequence of this method of synthesis and the non-reducing properties of isotrehalose are thus explained. Since the two C
atoms united by the
O linkage are
asymmetric, three stereoisomeric con-
figurations are possible for isotrehalose; to isotrehalose is uncertain.
which configuration belongs

is
The
disaccharide
upon
boiling with dilute mineral acids
hydro-
lyzed into d-glucose.
Dihexose Saccharides of Uncertain Nature and Constitution. In addition to the dihexose saccharides previously described a number of other sugars with the apparent formula C^HÔn have been reported by various investigators. Owing to the lack of confirmatory evidence
brief
mention
is
made
of only a
few of these compounds.
Astragalose reported by Frankforter* in the poisonous fruit of It reduces Fehling's solution and forms a Astragalus caryocarpus.
phenylhydrazone melting at 186 to 188 C. Parasaccharose formed according to Jodinf by the action of a
variety of Torula
It
upon sucrose
is
solutions in presence of
phosphate. duces Fehling's solution and

Pharbitose
[a]
forms
fine crystals, is dextrorotatory ([a]/
ammonium = + 108), re-
hydrolyzed by heating with acids.

in the seeds of Pharbitis Nil.
reported
by Kromerf
=+
109.53.
of
Pseudostrophanthobiose formed according to Feist in the hydrolysis pseudostrophanthin a glucoside occurring in Strophanthus hispidus.
*
Chem. Centralbl. Compt. rend., 53,
(1900) II, 484. 1252.
t Archiv.
Pharm., 234, 459.
Ber., 33, 2063, 2069.
730
Racefoliobiose reported
SUGAR ANALYSIS
by Boettinger
*
in grape leaves.
Revertose (Revertobiose)
of maltase or takadiastase
p. 704)
formed according to Hillf by the action
upon concentrated glucose solutions
(see
juice of Helix pomatia (the so-called

It is
Amygdalinbiose liberated according to GiajuJ by the action of the " edible snail ") upon amygdalin.
non-reducing and gives only d-glucose upon hydrolysis.
HEXOSE-HEPTOSE SACCHARIDES
XC
13
Galactosido-glucoheptose. This synthetic disaccharide was obtained by Fischer through reduction of the lactone of lactose carboxylic acid (p. 717) by means of sodium amalgam. The sugar, which has not been isolated in the
crystalline form,
is
dsHÔ^.
hydrolyzed by mineral acids into d-galactose and

2
d-glucoheptose.
CiH
4O M
Galactosido-glucoheptose
CH + H O = d-Galactose + CH
2
6 12 6
7
14
7.
d-Glucoheptose.
CiaHÔia. Glucosido-glucoheptose. This sugar was prepared by Fischer through reduction of the lactone of maltose carboxylic acid (p. 703) by sodium amalgam. The sugar, which has not been obtained in the crystalline form, gives
||
upon hydrolysis d-glucose and d-glucoheptose.

*
t
t
Chem. Ztg., 26, 24. Chem. News, 83, 578. Chem. Ztg., 34, 430.
Ber., 23, 937; Reinbrecht, Ann., 272, 197. Ber., 23, 937; Reinbrecht, Ann., 272, 197.
II
CHAPTER XXI
THE TRISACCHARIDES AND TETRASACCHARTOES
Trisaccharides
METHYLPENTOSE-HEXOSE SACCHARIDES
RHAMNINOSE.
,
(CH C 5 H 8 04)
3
>
(C 6 H n
5)
Occurrence and Preparation. Rhamninose is formed* by the the of xanthorhamnin glucoside hydrolysis by means of the enzyme which occurs associated with xanthorhamnin in Persian rhamninase,
berries (the fruit of
Rhamnus
infectoria).
prepared by extracting Persian berries with water; the enzyme is precipitated from the extract by means of alcohol. To obtain rhamninose xanthorhamnin is treated in aqueous solution beis
Rhamninase
tween 45 and 70 C. with a solution of rhamninase. The solution is then shaken out with ether to remove any unchanged xanthorhamnin, and then after clarification by means of bone black evaporated to a sirup; the sirup is extracted with hot alcohol, the alcohol solution evaporated The sugar is purified by to a sirup, and set aside for crystallization.
recrystallizing.
sists of
Rhamninose as above prepared conProperties and Reactions. white crystals with a composition and molecular weight corThe sugar has a mild, sweet responding to the formula Ci8H 32 Oi 4 fusion at 135 to shows 140 C. with decomposition incipient taste,
.
and hot alcohol, but not in ether. Rhamninose 41) and reduces Fehling's solution. levorotatory treatment with sodium amalgam rhamninose is reduced to Upon the alcohol rhamninite Ci8 H 34 Oi 4 ([a] D = - 57), which upon heating
and
is
is
soluble in water
= ([a] D
with dilute acids
is
hydrolyzed as follows:
2
= CH
6
14
12
5.
Rhamninite
Dulcite
Rhamnose.
is
Upon treatment with bromine

*
in
aqueous solution rhamninose
Tanret, Compt. rend., 129, 725. 731
732
oxidized
to
SUGAR ANALYSIS
rhamninotrionic
acid
CisHÔisCMu
94.3
is
for
acid-
lactone mixture), which
upon heating with

2
6
dilute acids
hydrolyzed as
5.
Ci 8
Rhamnino-
H O CH +2C H 2O + 2 H O = d-Galactonic Rhamnose.

3
15
12
12
trionic acid
acid
Upon oxidation with nitric acid rhamninose yields mucic acid. Upon warming with dilute hydrochloric or sulphuric acid rhamninose
is
hydrolyzed as follows: Ci 8 32 Oi4
+2H
= C
12
Rhamninose
d-Galactose
H +2C Rhamnose.
6
12
5.
These various reactions show that rhamninose is composed of 2 rhamnose and 1 d-galactose radicals, the latter having its aldehyde
group in a free reactive condition. Rhamninose is not fermented by yeast. Invertase, diastase and emulsin have no hydrolytic action. Rhamninose, through the presence of a reactive aldehyde group, forms a phenylhydrazone and osazone, but these compounds owing to their extreme solubility have not been obtained in a pure condition.
TRIHEXOSE SACCHARIDES / CeHiiOs
)C H
6
10
XC
RAFFINOSE.
Occurrence.
Melitriose.
HU
18
32
Gossypose. 5 2 0. 16
+ H
Raffinose
is
the best
known and most widely

raffinose
dis-
tributed of the trisaccharides.
The name
was
first
given to a
new sugar
discovered by Loiseau* in 1876 in the impure molasses
obtained from refining beet sugar (French, raffiner
to refine).
The
as
same sugar had been previously

Eucalyptus
melitose.
isolated,
however, by Johnstonf from

described
manna
in
1843, afterwards
by Berthelott
Tollens, however, showed that melitose was identical with Loiseau's raffinose and also proved the same to be true of gossypose, a sugar found by Ritthausen and by Bohm 1f in cottonseed meal. The identity, thus established by Tollens, was important for it opened the way to investigations which established the wide occurrence of
||
raffinose in the vegetable
mentioned
*
raffinose has
kingdom. In addition to the sources just been found in barley and other grains, in young
Ber., 18, 26.
II
J. fabr. sucre., 24, 52; 26, 22.
t J. prakt. t
Ann.
29, 485. chim. phys. [3], 46, 66.

[1],
Chem.
J.
prakt.
If
J.
prakt.
Chem. Chem.
[2],
[2],
29, 351. 30, 37.
THE TRISACCHARIDES AND TETRASACCHARIDES

wheat sprouts (up to
733
6.9 per cent of the dry substance) and in many other plant substances. Raffinose has attracted most attention from its occurrence in sugarbeet products. It had been the opinion of many chemists that raffinose
was formed during the process of manufacture by the action of alkalies upon the sucrose, invert-sugar and other constituents of the juice; Lippmann,* however, was able to separate raffinose directly from expressed beet juice, thus proving that the sugar was formed during the growth of the beet. The amount ordinarily occurring in sugar beets is only from 0.01 per cent to 0.02 per cent; under certain conditions,
however, the percentage of raffinose
sult,
may greatly exceed this, the reperhaps, of abnormal climatic causes, such as drought, excessive rain, freezing, etc., the exact role of these various factors being as yet not clearly understood. The synthesis of raffinose in the plant is apparently connected with a saccharification of galactan substances (pectin, etc.) in presence of sucrose, the result no doubt of enzyme
action.
Raffinose has also been reported
by
Pelletf
in sugar-cane molasses, although the claims for this
and other investigators have been dis-
Until additional evidence is obtained the by Lippmann. J occurrence of raffinose in sugar-cane products must be regarded as exceedingly unusual.
puted
Raffinose may be prepared from Preparation of Raffinose. Eucalyptus manna (a secretion from certain Eucalyptus trees, as E. viminalis and E. Gunnii) by extracting the manna with hot water, clarifying the extract with bone black and evaporating to the point of
more common material for preparing raffinose is cottonseed meal; the method of Zitkowski is as follows: Cotton-seed meal Preparation of Raffinose from Cottonseed Meal. is extracted with cold water for 1 hour and the filtered extract made
crystallization.
faintly alkaline with milk of lime. is polarized (in terms of sucrose)
The filtrate from precipitated matter and then treated at low temperature
with finely powdered quick lime in the proportion of 125 parts CaO to 100 parts of sugar. The precipitate of calcium raffinosate is filtered off,
washed with cold lime water and then,
after dissolving in hot water, carbonated at 80 C. almost to neutrality. After filtering from calcium carbonate the solution is concentrated to a sirup of about 75 degrees Brix and set aside in the cold to crystallize. Priming with a crystal of raffi-
nose will hasten the process.

*
The
crystals of raffinose are filtered off,

t
Ber., 18, 3087.
t Bull, assoc.
chim. sucr.
diet., 14,
139.
Deut. Zuckerind., 22, 1439. Am. Sugar Ind., 12, 324.
734
SUGAR ANALYSIS
washed with 90 per cent alcohol and purified by recrystallization. In one experiment by this method 600 gms. of raffinose hydrate were obtained from 150 Ibs. of cottonseed meal.
Preparation of Raffinose from Beet Molasses. haye been devised for preparing raffinose
A number of processes
from beet molasses. Scheibler* first noted that absolute methyl alcohol had a high solvent action upon raffinose (9.8 gms. raffinose anhydride in 100 c.c.) and a
low solvent action upon sucrose (0.4 gm. sucrose in 100 c.c.). Using this observation as a basis Burkhardf employed the following method A low-grade beet molasses rich in raffinose (preferably from the strontium monosaccharate process) is absorbed upon clean dry wood shavings and, after thoroughly drying in a vacuum, extracted with absolute
:
methyl alcohol. The extract is diluted with water, the alcohol evaporated and the solution boiled, during addition of crystallized strontium hydrate with constant stirring, until a permanent crust of crystals beThe strontium compound is filtered off, gins to form upon the surface. washed with hot saturated strontium hydroxide solution and then carbonated in suspension with water. The solution is evaporated, the sirup dissolved at 60 to 70 C. in the exactly necessary amount of 80 per cent alcohol, and then set aside for 24 to 48 hours, when raffinose will
crystallize out.
The precipitation of raffinose from molasses as lead raffinosate by means of ammoniacal lead acetate, lead carbonate or litharge has also
been successfully employed. ZitkowsldJ has used the following process, which is based upon the insolubility of lead, raffinosate and the
solubility of lead saccharate at high temperature: Thirty pounds of the molasses are diluted to
about 50 degrees
Brix, brought to a boil with the addition of 3 pounds of litharge and filtered, this being done for the purpose of precipitating some of the lead salts that form. Then 3 pounds more of lead oxide are taken and
just sufficient of the purified molasses filtrate added to form a thin The mixture is stirred for about an hour in the cold when the paste.
formation of lead saccharate begins; the mass which becomes stiff is then allowed to set twenty-four hours. The main portion of the molasses
solution
is
then brought to a boil and the lead saccharate added in small
When all of the portions at a time in order to disintegrate the mass. lead saccharate is added, the mixture is kept at boiling for about thirty minutes, then filtered and thoroughly washed with water. The lead com* t
Ber., 19, 2868. Neue Ztschr. Rubenzuckerind., 20, 16.
Am. Sugar
Ind., 13, 8.

pound thus obtained
735
is decomposed with carbon dioxide, filtered and evaporated to a light sirup. The sirup is treated with blood black and again filtered, evaporated on a water bath to a heavy sirup and set away to crystallize. The filtration of the lead raffinosate should be performed as hot and as quickly as possible, otherwise considerable
quantities of lead saccharate will be precipitated.

of the final sirup can be accelerated
raffinose.
The crystallization by priming with a pinch of pure
Raffinose crystallizes from aqueous solution in the Properties. form of long pointed needles or prisms, with a composition and molecular weight corresponding to the formula Ci 8 H 32 Oi6 + 5 H 2 O. The crystals upon gradual warming at 80 C. for several hours and then at 100 to 105 C. lose all their water and pass without melting into the anhydride. Upon rapid heating the crystals melt in their water of crystallization below 100 C.; under this condition the last traces of water are removed only at 125 to 130 C. when decomposition sets in with brown coloration and odor of caramel. The sensibility of raffinose
to destructive changes
upon rapid heating is shown by raffinose-containing beet sugar, which darkens at 120 to 125 C.; while ordinary beet sugar, free from raffinose, is not as a rule affected.
Raffinose anhydride has the formula Ci8H 32 Oi6 and consists of a white amorphous hygroscopic mass which upon exposure to moist air reabsorbs after several days the entire amount of water of crystallization.
Raffinose hydrate is more soluble than sucrose in hot water, but less soluble in cold water; 14 to 15 parts of water are necessary to dissolve raffinose at SupersatuC., 9 parts at 10 C. and 6 parts at 16 C.
rated solutions are easily formed from which the raffinose is deposited upon long standing. Raffinose is insoluble in absolute ethyl. alcohol or
in ether;
its solubility
in absolute
methyl alcohol
is
considerable as
previously stated. Raffinose, through its property of combining with water of hydration, seems to possess the property of throwing sucrose out of solution.*
The Influence of Raffinose Upon the Crystalline Form of Sucrose. of sucrose in of effect raffinose exerts a giving crystals presence peculiar a pointed needle-like structure; 3 per cent raffinose in a sugar sirup may
produce a sensible elongation of the grain, the pointed character of the This alteracrystals increasing with the amount of raffinose present. tion in grain is frequently noted in the crystallization of low-grade beet products and is usually an indication of the presence of raffinose.
*
Herzfeld, Z. Ver. Deut. Zuckerind., 42, 207.
736
It
SUGAR ANALYSIS
may
when
produce under certain concrystallization takes place
must be remembered, however, that other impurities (organic lime
caramelization products, etc.) ditions a pointed grain, especially

salts,
the other hand, raw beet from viscous supersaturated solutions. of raffinose without alteration of 4 cent to 5 per sugars may contain in molasses coating of the remains dissolved grain, in case the raffinose
the crystals.*
Specific Rotation.
On
In aqueous solution raffinose
is
strongly dextro-
104 to 105.7, rotatory, the value of [a] D for the hydrate ranging from different for observers different to preparations of sugar. according
For purposes of analysis the value
104.5
may
be used without serious

'
.
(104
100J-
The observations
Reactions.
of
Creydtf show a slight
falling off in specific rota-
tion with increase in temperature.

in this respect similar to sucrose.
having
Raffinose does not reduce Fehling's solution, beRaffinose also shows the
same
resistance to the action of alkalies as sucrose.
Both
of these re-
actions indicate the absence in raffinose of a functional aldehyde or ketone group. Upon oxidation with nitric acid raffinose yields a mixture
of acids, of which oxalic, saccharic and mucic are the most important. The yield of mucic acid from raffinose hydrate by the method of Tollens
is
22 to 23 per cent, which corresponds to about 30 per cent galactose (yield of mucic acid from galactose by same method is 77 to 78 per
cent).
Hydrolysis of Raffinose by means of Acids. Upon heating with dilute hydrochloric or sulphuric acid raffinose is hydrolyzed according to
the following equation:
18
32
16
+2H
= CH
6
12
Raffinose
d-Glucose
H + +C d-Galactose
6 12 6
CH
6
12
6.
d-Fructose.
The total yield of reducing sugars according to this equation would be 107.1 per cent for the anhydride and 90.9 per cent for the hydrate. The
yield of galactose from raffinose hydrate according to theory would be 30.3 per cent, which agrees closely with the value calculated from the yield of mucic a'cid.
eral phases.
The hydrolysis of raffinose, as Tollens J first showed, proceeds in sevThe first step in the reaction is the splitting off of fructose;
glucose and galactose appear only at a later stage of the reaction. Scheibler and Mittelmeier showed that by moderate warming with
*
t Z. Ver.
Herzfeld, Z. Ver. Deut. Zuckerind, 39, 661. Deut. Zuckerind. 37, 153.
Ann., 232, 169.

Ber., 22, 1678, 26, 2930.

dilute acid (as 10 gms. raffinose
737
+ 90 c.c. water + 6
+
d-Fruotose
acid 1.19 sp. gr. 10 minutes at
68) the
c.c. hydrochloric reaction proceeds as follows:
CisHÔie
Raffinose
-f-
HÔ = CeHÔe
C^HÔu.
Melibiose.
It is only by prolonged heating with more concentrated acid that the melibiose (see p. 723) is hydrolyzed into d-glucose and d-galactose, the complete conversion of the melibiose being accompanied by a partial
destruction of the fructose.
As a
is
rule less
theoretical yield of monosaccharides

of raffinose
than 90 per cent of the obtained by the acid hydrolysis
under the most favorable conditions. During the hydrolysis of raffinose the specific rotation undergoes a marked decrease, the final reading depending upon the extent of the hyFor the ordinary method of Clerget inversion the specific rotadrolysis. tion of raffinose hydrate decreases from + 104.5 to about + 53 or + 54, which corresponds to the mixture of fructose and melibiose required by the preceding equation (30.30 per cent fructose and 57.57 per cent melibiose).*
raffinose
Upon prolonged
heating with acid the specific rotation of

+- 20.
was found by Tollens to sink as low as

is
The
theoretical
value f for a mixture of 30.3 per cent each of d-glucose, d-galactose and
d-fructose
about
12.50;
decomposition of fructose, however, sets
in before this limit is reached so that higher figures of variable value
are obtained.
raffinose
The hydrolysis of Means of Enzymes. can also be effected by means of enzymes, the nature of the reaction depending upon the character of the enzyme.
Hydrolysis of Raffinose by
Invertase hydrolyzes raffinose into d-fructose and melibiose, as already described under the latter sugar. Emulsin, on the other hand, hydrolyzes raffinose into d-galactose and sucrose. The general formula for both of these reactions is the same
:
C H 20 + H
18
3 16
/
I
= CH
= =
6
12
T?
invertase
4-
emulsin
d-fructose d-galactose
++CuHaOii.
melibiose.
+ sucrose.
is
For the complete hydrolysis of raffinose into saccharides the action of two different enzymes
*
its
component mononecessary and

it is
The 57.57 per cent
= +15.9; 30.3 per cent d-glucose would give a rotation of 0.303 X +52.5 = the 30.3 X 0.303 would +81 per cent the 30.3 per cent d-galactose +24.5; give The combination of these effects 27.9. 92 = d-fructose would give 0.303 X
t
X +143 = +82.3; the 30.30 = -27.9. The combination

The
melibiose anhydride would give a rotation of 0.5757 92 per cent fructose would give a rotation of 0.303 X 27.9 = +54.4. of those effects would be +82.3
would be +15.9
+ 24.5 -
27.9
=+12.5.
738
SUGAR ANALYSIS
evident that this can be accomplished by the action of invertase and melibiase (p. 723), or by that of emulsin followed by invertase. These reactions may be explained by assuming the following ar-
rangement
for the
monosaccharide groups in
t
6
10
raffinose.
C H n O5 - O - C H O 4 - O - C Hu0
6 6
5.
d-Fructose
radical
d-Glucose
radical
d-Galactose
radical
Sucrose
Melibiose.
The hydrolysis by means of invertase or weak acids takes place at O atom marked *; the hydrolysis by means of emulsin takes place atom marked fat the The fermentation of raffinose by Fermentation of Raffinose. means of yeast depends upon the character of the enzymes which are Bottom yeasts, which contain both invertase and melibiase present.
the
and can thus effect a complete hydrolysis, ferment raffinose completely, although somewhat more slowly than sucrose. Top yeasts, on the other hand, which do not ordinarily contain melibiase, ferment only the
fructose part of the molecule with a corresponding reduction in the The theoretical equations for the two fermentations yield of alcohol.
would be:
Bottom
yeast,
Ci 8
32
Oi 6 +2
2
HO =
2
C
5
H OH
5
Raffinose
Alcohol (54.78 per cent)
6 C0 + Carbon
2
2.
dioxide.
Top
yeast, Ci 8
32
Oi 6
+HO =
C H OH
2
Raffinose
Alcohol (18. 2 6 per cent)
CO + +2 Carbon
dioxide
Ci 2
22
On.
Melibiose.
The yield of alcohol, expressed in percentage of raffinose anhydride, somewhat less in actual practice than indicated above. A number of moulds (species of Monilia, Amylomyces and AspergilOther moulds, as Aspergillus lus) also hydrolyze and ferment raffinose. niger and Penicillium glaucum, hydrolyze raffinose but instead of producing alcohol form acid oxidation products such as oxalic and succinic
is
acids.
Special bacteria also ferment raffinose with production of lactic and butyric acids. Compounds of Raffinose. Owing to the absence of a reactive
aldehyde or ketone group, raffinose does not form hydrazones, osazones, mercaptals, ureides, oximes or any other of the numerous compounds
which are characteristic
of reducing sugars. heating raffinose with acetic anhydride Scheibler and Mittelmeier* obtained a hendecacetate, Ci 8 H2i(C 2 3 O)iiOi6. After recrys-
Upon
Ber., 23, 1438.

tallizing
from hot absolute
alcohol, the
compound was obtained
as
white
melting at 99 to 101 C.; it is soluble in absolute alcohol, 92.2. aniline, chloroform and benzol and is dextrorotatory, [a] D = Tanret* has prepared a dodecacetate of raffinose, Ci 8 20 (C 2 3 0)i 2 Oi 6
leaflets
[a] D
=+100.3.
S toilet obtained by the usual methods a raffinose octobenzoate, 24 (C7H 5 0) 8 Oi6; the compound consists of a white powder, melting at 98 C. and showing in glacial acetic acid [oi\ D = -f- 4.1. Raffinose forms a number of compounds with the alkalies, alkaline earths and other metals. These compounds have been especially studied by Beythien and TollensJ from whose work the following exCi8
amples are taken.
an alcosodium alcoholate. By taking a two-molecular proportion of sodium alcoholate the compound Ci 8 H 3 iNaOi6 + NaOH is obtained. Both substances are
raffinosate,
Sodium
Ci 8
H iNaOi6, is obtained
3
by
precipitating
holic raffinose solution with a one-molecular proportion of
white amorphous powders.
Barium raffinosates, corresponding to the formulae Ci8 32 Oi 6 'BaO and Ci 8 H 32 Oi62BaO, are obtained by mixing barium hydroxide and raffinose solutions in presence of alcohol in the proper molecular proportions.
The compounds were obtained
as
white amorphous substances of
imperfect purity. Strontium raffinosate, of the formula Ci8 32 Oi 6 2 SrO 2 0, is obtained by heating a solution of strontium hydroxide and raffinose, as a sticky mass which becomes granular upon long boiling or upon addi-
+H
The compound consists of a white granular amorphous powder which loses its water of combination at 80 C. Raffinose compounds containing 1 SrO and 3 SrO have not as yet been obtained. Calcium raffinosate, Ci 8 H 32 Oi6 '3 CaO + 3 H 2 O, is obtained by heating
tion of alcohol.
a raffinose solution saturated with calcium hydroxide. The compound consists of a white amorphous powder which loses its water of combination at 100 C.
by dissolving calcium hydroxide in a cold raffinose solution obtained the compound Ci8 H 32 Oi 6 -2 CaO + 5 H 2 O. Lindet also noted upon treating a solution of sucrose, raffinose and lime with alcohol that calcium raffinosate was dissolved mostly by weak and calcium saccharate mostly by strong alcohol. This method has been proposed as a means of separating sucrose and raffinose, but is inferior to the methods deLindet
scribed under preparation of raffinose.
* Bull. soc. chim.
[3],
13, 261.
t Z.
Ver. Deut. Zuckerind., 39, 894.
f Z. Ver. Deut. Zuckerind., 61, 33.
J. fabr. sucre, 31, 19.
740
Lead[raffinosate,
SUGAR ANALYSIS
Ci 8
H 2Oi
3
6 '3
by heating raffinose solutions with litharge or, according to Wohl,f more advantageously by heating with lead saccharate (see under preparation of raffinose). As in the case of most other sugars the Tests for Raffinose.
only absolute test for raffinose is the separation of the sugar in pure crystalline form and the determination of its specific rotation, products For the separation of raffinose of hydrolysis and other properties.
upon Lead
treating raffinose solutions with raffinosate can also be prepared
PbO, was obtained by Lippmann* ammoniacal lead subacetate.
any
will
of the
It is evident
methods described under preparation may be used. from its composition that raffinose after hydrolysis the reactions described for d-glucose, d-fructose and of give any
d-galactose, so that ordinary qualitative tests are valueless when The removal of fermentable sugars several of these sugars are present. by a pure culture of top yeast, and examination of the residual sugars
for melebiose
may be used for
corroboration.
For quantitative methods
of determining raffinose see page 281.
The probable arrangement of the monosaccharide Configuration. groups in raffinose has already been given; the manner in which these different groups are combined has not, however, been established.
The
following configuration is regarded at present as the one which corresponds most closely to the properties of raffinose.
CH OH
2
d-Galactose radical
d-Glucose radical
d-Fructose radical
The
synthesis of raffinose has not as yet been effected.
Melezitriose. 2 H2 O. Ci 8 H32Oi 6 Occurrence. This trisaccharide, first observed in 1833 by Bonastre,J has been found for the most part as a constituent of the secre-
MELEZITOSE.
manna of Pinus larix, manna of Alhagi Maurorum (Turkestan manna), Lahore manna, honey dew of the
tions of different trees, such as
*
t
j
Z. Ver. Deut. Zuckerind., 36, 257. Deut. Zuckerind., 25, 1125. J. pharm. chim. [2], 8, 335; 19, 443, 626.
741
The sugar was named melezitose by Berthelot* in 1856, linden, etc. and was supposed by him to be a disaccharide; Alekhine,f however,
proved the sugar to be without question a trisaccharide. For the preparation t of melezitose Preparation.
manna
(Turandjabine)
is
extracted with
solution concentrated to a sirup;
Turkestan and the filtered water, an excess of methyl alcohol is then
warm
added when
is
crystallization takes place within 24 hours.
The crude
bone black; coloring matter is precipitated purified sugar little barium a hydroxide solution, any excess of the latter being reby moved with ammonium carbonate. The filtrate is again concentrated and The yield of pure melezitose crystallized in presence of methyl alcohol.
of
by means
36 per cent of the manna taken. Melezitose as ordinarily prepared consists of white Properties. rhombic crystals with a composition and molecular weight correspond2 2 O. The crystals of the hydrate ing to the formula Ci 8 H 3 20i 6
by
this
method
is
+ H
effloresce
upon exposure to the air and with gradual elevation of temup their water, passing without decomposition into the give perature The latter may also be obtained directly upon anhydride Ci8H 32 Oi 6
.
crystallizing melezitose
solutions.
from hot concentrated aqueous or alcoholic
Melezitose anhydride consists of a white crystalline powder which upon rapid heating melts at 148 to 150 C.; it is soluble in 2.73
parts of water at 17.5 C. and 0.32 part at 100 in hot alcohol but insoluble in ether.
C., it
is
slightly soluble
Melezitose
for the
is
dextrorotatory,
83. hydrate Hot solutions of dilute alkalies are Reactions and Hydrolysis. without action upon melezitose. The sugar like raffinose does not reduce Upon heating with dilute hydrochloric or sulphuric Fehling's solution.
anhydride Mutarotation does not exist.

[a] D
for the
=+
88.5
and
acid melezitose
is
hydrolyzed, the reaction proceeding as Alekhinell
found in two distinct stages. The first phase of the hydrolysis consists in the conversion of melezitose into d-glucose
and the disaccharide turanose.
Ci 8
32 Oi6
+HO
2
= CH O
6 12
Ci 2
22
On.
(1)
Melezitose
d-Glucose
is
Turanose.
best performed by means of 20 per cent hydrochloric acid in the cold or upon warming with 1 per cent sulphuric
This part of the hydrolysis

*
Compt.
rend., 47, 224.
t Bull. soc. t
chim.
"
[2],
46, 824.
Maquenne's Les Sucres." p. 701. Turkestan manna, or Turandjabine, is used in the Orient for sweetening = sand; It is sold in Tashkend under the name of Koum-tchakar (Koum drinks. 532. chim. Ann. = 18, phys. [6], tchakar sugar).
II
742
SUGAR ANALYSIS
83 for the hydrate to about acid; the rotation of the sugar falls from first of the the marks 63 which -f step in the hydrolysis. completion dilute with hydrochloric or sulphuric acid, Up6n prolonged boiling
melezitose
is
completely hydrolyzed into d-glucose
CigHwOie
Melezitose
+2H
CH
6
12
6.
(2)
d-Glucose.
In this second phase of the hydrolysis the turanose, which is first formed, is split up into two molecules of d-glucose, and the specific
rotation falls to about
d-glucose.
+51
which agrees very closely with that of
In the second stage of the hydrolysis, according to Tanret, turanose hydrolyzed into d-glucose and d-fructose, so that melezitose would give upon complete hydrolysis 2 molecules of d-glucose and 1 molecule of
is
d-fructose.
The
calculated rotation of the latter mixture would be
about
for
+ 4.5 which
does not agree with the value obtained by Alekhine

is
hydrolyzed melezitose. Additional investigation cide the question. (See under turanose, p. 725).
needed to de-
Upon acetylating with acetic anhydride Alekhine Compounds. obtained a hendecacetate, CisH 2 i(C 2 3 0)iiOi6, which consists of large monoclinic prisms melting at 170 C.; it is non-reducing, insoluble in
water, soluble in alcohol and benzol and shows in benzol solution
[^ = +110.44.
Fermentation.
gillus niger effects
ose,
but
is
Melezitose is not fermented by yeast. Aspera slow hydrolysis at 50 C. into d-glucose and turanwithout further change.
Ci 8
GENTIANOSE.
Occurrence.
H 2Oi
3
6.
This trisaccharide was discovered by Meyer* in the roots of Gentiana lutea and, according to Bourquelot and Nardin,f occurs also in other members of the Gentian family. Fresh Gentian roots are ground and extracted for Preparation. 20 to 25 minutes with boiling 95 per cent alcohol using a reflux condenser.
The
alcoholic extract
is
pressed out, the alcohol distilled
off,
excess of calcium carbonate
added to neutralize acid and the solution filtered. The filtrate is evaporated to a sirup, freed from gummy matter by precipitation with 95 per cent alcohol, and the clear alcoholic solution filtered and set aside. Crystals of gentianose separate after about 2 weeks' standing; they are filtered off and purified by recrystallization. The Gentian roots employed for the preparation of gentianose must
be fresh; old or dried roots or aqueous extracts do not yield gentianose
*
Ber., 15, 530; Z. physiol.
Chem.,
6, 135.
Compt.
rend., 126, 280.

on account of
obiose.
its
743
hydrolysis
by an enzyme
is
into d-fructose
and genti-
C. and having a composition and molecular to the formula C 18 H32O 16 The sugar is easily weight corresponding soluble in cold water, slightly soluble in boiling alcohol, insoluble in
to 210
.
Properties. melting at 209
Gentianose
obtained in the form of white crystals,
absolute alcohol and ether.
Gentianose
is
dextrorotatory,
[a]
31.25 (Bourquelot and Nardin). After boiling the solution (Meyer), noted in one instance [a] D = 65.7. Whether this is due to Meyer
+ 33.4
mutarotation or to some chemical change does not reduce Fehling's solution.

Hydrolysis of Gentianose.
is
uncertain.
Gentianose
Gentianose upon heating with acids undergoes hydrolysis, the reaction proceeding as shown by Bourquelot and Herissey in two distinct stages. In the first phase of the hydrolysis gentianose is hydrolyzed into d-fructose and the disaccharide gentiobiose
(p. 726).
CisHÔie Gentianose
obiose.
+ H
=
is
CeHÔe
d-Fructose
Ci 2
22
On.
Gentiobiose.
This step of the hydrolysis
best carried out as described under genti-
Upon
(p. 726). follows
:
obiose which
heating gentianose with 3 per cent sulphuric acid, the gentiis first split off is hydrolyzed into 2 molecules of d-glucose
The complete
hydrolysis of gentianose
2
is
then expressed as
6
C H
18
32
16
+ 2H
CH
6
12
+ 2C H
12
Gentianose
d-Fructose
d-Glucose.
Fermentation and Action of Enzymes.

third fermented
Gentianose
is
only one-
by
yeast, the invertase of the latter splitting off d-fruc-
tose, which is fermented, and the gentiobiose remaining unfermented. Aspergillus niger contains enzymes, which effect the complete hydrolysis
of gentianose,
and thus ferment the sugar entirely. Diastase and emulsin are without action on gentianose. Emulsin, however, can hydrolyze gentiobiose, so that yeast in presence of emulsin can ferment
gentianose completely. According to Bourquelot emulsin seems to be accompanied at times by an enzyme which hydrolyzes gentianose into
d-glucose and sucrose.
Configuration.
in gentianose
is
The arrangement
of the
monosaccharide groups
probably as follows:
CH,,05 - O - C6 H 10
d-Fructose
radical
t - O - C HU
6
6.
d-Glucose
radical
d-Glucose
radical
Sucrose
Gentiobiose
744
SUGAR ANALYSIS
hydrolysis by means of weak acids or invertase takes place at atom marked *. The hydrolysis into d-glucose and sucrose would take place at the O atom marked f- The non-reducing properties of gentianose show that none of its monosaccharide components contains a reactive aldehyde or ketone group; the manner in which the
The
the
monosaccharide groups of gentianose are united is not known, so that the configuration of this trisaccharide still remains uncertain.
MANNATRISACCHARIDE,
Occurrence.
in the
Ci 8
32
Oi 6
Mannatrisaccharide was discovered by Tanret
manna
it
which
makes up from about 6
of the ash tree (Fraxinus Ornus, F. rotundifolia, etc.), of to 16 per cent. Ash manna also
contains from 40 to 60 per cent of mannite and a smaller amount of mannatetrasaccharide or stachyose (see p. 747) in the preparation of mannatrisaccharide it is necessary to remove these accompanying con;
stituents
by
fractional crystallization
and
precipitation.
Ash manna is extracted with 70 per cent alcohol, Preparation. and the mannite which crystallizes out separated by filtration. The mother liquor is then evaporated to dryness and extracted first with
and then with boiling 85 per cent alcohol. In this is mostly eliminated and a residue obtained showThe solution of the residue is ing a rotation of about [a] D = + 140. then fractionally precipitated with barium hydroxide in presence of alcohol; the two fractions are decomposed separately with carbon dioxide to precipitate barium and the solutions evaporated io crystallizaboiling 95 per cent
manner the mannite
tion.
The crude sugars

is
trisaccharide
are recrystallized several times when mannaobtained from one portion and mannatetrasaccharide
from the other.

Mannatrisaccharide is a white sweet crystalline subProperties. It is easily stance, very hygroscopic and melting at about 150 C. soluble in water, soluble at 15 C. in 60 parts 85 per cent and in 130
parts 90 per cent alcohol and at 78 C. in 200 parts absolute alcohol. Mannatrisaccharide reduces Fehling's solution about one-third as strong as d-glucose and is strongly dextrorotatory, [a] D = 167.
Upon
into 1
heating with dilute acids mannatrisaccharide is hydrolyzed molecule of d-glucose and 2 molecules of d-galactose.
C H O
18
32
16
+ 2H O
2
CH O
6 12
Mannatrisaccharide
+ 2C H O
6
12
d-Glucose
d-Galactose.
Upon oxidation with bromine in aqueous solution mannatrisaccharide

*
Compt.
rend., 134, 1586.

is
745
dilute acids
oxidized to mannatrionic acid, C^HÔn, which is hydrolyzed as follows

:
upon warming with

6
18
32
17
+ 2H
= CH
6
12
Mannatrionic acid
+ 2C H
12
d-Gluconic
acid
d-Galactose.
This reaction shows that the functional aldehyde group of mannatrisaccharide belongs to the d-glucose group.
Mannatrisaccharide is slowly fermented by yeast, but the completeness of this fermentation has not been determined.
Owing to the presence of a reactive aldehyde group mannatrisaccharide forms with phenylhydrazine a yellow amorphous hydrazone, easily soluble in water and alcohol, and a crystalline osazone melting
at 122
C. and quite soluble in water.
Mannatrisaccharide forms a dodecacetate, Ci8H 2o(C 2 3 O)i 2 Oi6, ([a] D in alcohol = +135). Barium hydroxide, in presence of alcohol, precipitates Ci 8 H 32 Oi 6 BaO and ammoniacal lead subacetate, Ci8H 24 Pb 4 Oi6.
Lactosin. Ci 8 H 32 Oi 6 ? This sugar was discovered by Meyer* in the roots of Silene vulgaris and other Cariophyllacese. It has also been found in
Lactosinose.
Occurrence.
Quillai-bark (bark of Quillaia Saponaria) and in Saponaria rubra. Preparation. The expressed juice of the roots of Silene vulgaris
is
treated with an excess of strong alcohol. The precipitate is dissolved in water, clarified with lead subacetate, the solution filtered and treated
with ammoniacal lead acetate; the lead lactosinate is filtered off, decomposed in aqueous suspension with hydrogen sulphide, the solution
filtered from lead sulphide and evaporated; the sirup thus obtained is treated with strong alcohol and the precipitated sugar, consisting of an amorphous mass, dried first over concentrated sulphuric acid and then at
110 C.
The
dried product
is
then boiled
to 3 days with 80 per cent
alcohol under a reflux condenser; the quantity of alcohol should not be sufficient to dissolve all the crude sugar. Upon filtering the alcoholic
extract
and
cooling, the lactosinose

if
is
deposited as crystals, which
may
be purified
necessary by recrystallization.
Lactosinose, as above prepared, consists of small glistening crystals which, after drying over concentrated sulphuric acid, have a composition corresponding to Ci8H 32 Oi 6 ( or C 36 6 4O 32 ). The sugar is
Properties.
easily soluble in water, somewhat soluble in 50 per cent alcohol and is The concentrated dissolved by 350 parts of hot 80 per cent alcohol.
very viscous. Lactosinose is strongly dextroUpon drying at 110 C. lactosinose berotatory, [a]^=+211.7.
aqueous solution
is
Ber., 17, 685.
746
comes amorphous and a D = + 168 to + 190.
[
SUGAR ANALYSIS
in
this
condition
shows a lower
rotation,
not affected by boiling solutions of dilute alkalies and does not reduce Fehling's solution except after long boiling (7 minutes) when a very slight reduction may take place. Upon oxidation with nitric acid a large amount of mucic acid is formed. Upon
Lactosinose
is
boiling a 1 per cent solution of the sugar with hydrochloric or sulphuric acid, using 1 part acid to 1 part sugar, lactosinose is slowly hydrolyzed,
The products of the 50. the specific rotation diminishing to below cent 45 consist of about d-galactose the presence of an per hydrolysis
;
undetermined dextrorotatory and of an undetermined levorotatory sugar

is
also indicated.
The compounds and other properties of More study is required upon investigated.
stitution
lactosinose have not been

lactosinose before its con-
and
its
exact relationship to other carbohydrates can be
tabulated.
Secalose.
Occurrence.
/3-Levulin.
CigHÂe.
Secalose, formerly called jS-levulin, was discovered by Schulze and Frankfurt* in green rye (Secale cereale), where it occurs to the extent of 2 to 3 per cent. It has also been found in green oats
in ray-grass (Lolium perenne). The alcoholic extract of green rye, or oats, is treated Preparation. with strontium hydroxide solution and the strontium secalate, which is
and
precipitated, filtered off, decomposed with carbon dioxide
and the secalose
precipitated from the evaporated filtrate by means of strong alcohol. Purification of the sugar is carried out in the same manner as described
for stachyose.
Secalose crystallizes as a hydrate in the form of Properties. white microscopic prisms, which upon heating in a stream of dry hydrogen at 100 C. lose all their water without decomposition. The
anhydrous sugar has a composition corresponding to the formula The sugar is easily soluble in water, in which it exhibits Ci 8 H 3 20i 6 28.6 to 31.7. It does not reduce Fehling's levorotation, [a] D =
.
solution.
Secalose upon warming with dilute hydrochloric or sulphuric acid is No other sugar has been detected rapidly hydrolyzed into d-fructose. among the products of hydrolysis. Additional investigation is required upon secalose before its constitution
and
its
relationship to other sugars can be determined.

*
Ber. 27, 65, 3525.

The Tetrasaccharides TETRAHEXOSE SACCHARIDES
/
747
) \ X C H 11
6 5
STACHYOSE.
Occurrence.
ash
Mannatetrasaccharide.
C24H 42 02i
+ 4 H 0.
2
The discovery
of a tetrasaccharide
by Tanret
in
has already been mentioned (p. 744). Tanret* showed later that this tetrasaccharide was identical with a sugar found by Plantaf
in the tubers of Stachys tuberifera
manna
and named by him stachyose.
The
sugar has also been found in the roots of different plants belonging to the Labiatse, in the roots of Lansium altuus and in the white jasmine.
Preparation.
Stachyose
according
to
Schulze
and
PlantaJ
makes up from 14.16

of Stachys tuberifera.
to 73.07 per cent of the dry substance of the tubers

is clarified
To prepare the sugar the expressed juice of the tubers
with
lead subacetate and mercuric nitrate, the lead and mercury are precipitated from the filtrate by hydrogen sulphide and the clear solution neutralized with
ammonium
is
hydroxide, and evaporated to a sirup.
The
sirup thus prepared
poured into
an excess of alcohol which throws down
an abundant
precipitate.
The
latter is separated, dissolved in a little
water, clarified
with phosphotungstic acid, filtered, the excess of clarifying removed with barium hydroxide solution, again filtered and satuagent rated with carbon dioxide to remove barium; the barium carbonate is
filtered off,
the filtrate concentrated and again poured into alcohol which
The stachyose is purified by precipitates flakes of impure stachyose. in water and the flakes precipitating with alcohol, repeating dissolving the process several times; the product is finally dissolved in a little
water, alcohol added
for crystallization,
little
till
the strength of the solution

filtered off
is
91 per cent;
any
If
precipitated stachyose
is
and saved and the
filtrate set aside
which usually requires several weeks' standing.

is
crystallized stachyose be hastened by priming. Stachyose is obtained as hard rhombic crystals of Properties. sweet taste and with a composition corresponding to the formula 4 2 O. The water of crystallization is partially removed
available the process of crystallization
may
+ H
Compt.
rend., 136, 1569.
Landw. Vers. Stationen,
25, 473.
| Ber., 23, 1692; 24, 2705.
748
SUGAR ANALYSIS
upon standing over concentrated sulphuric acid or upon warming to 100 C. The water is completely removed at 115 to 120 C. decomposition and oxidation set in, however, below this temperature so that the anhydride cannot be prepared in this way. The best method of de;
hydration is to heat the powdered sugar in a stream of dry hydrogen at 103 C. for half an hour; in this manner all water is removed without decomposition.
Stachyose
dissolved
is
easily soluble in water, 1 part of the hydrate being
by
in 14 parts
C.; at 15 C. the hydrate is soluble 60 per cent, in 55 parts 70 per cent and in 300 parts 80 per
It is insoluble in absolute alcohol.
0.75 parts water at 13
cent alcohol.
is strongly dextrorotatory, [a] D for the anhydride +148.1 (Schulze and Planta) and +148.9 (Tanret); [a] D for the hydrate 132.75 to +133.85 (Tanret) and +133.5 (Schulze). If +148.5 be taken for the anhydride, the theoretical [a] for C24H42021 D
Stachyose
to
= +147.9
=+
+4HO
2
is
134.0.
Stachyose is not affected upon heating with dilute solutions of alkalies and does not reduce Fehling's solution. Upon oxidation with
nitric acid
stachyose yields 37 to 38 per cent mucic acid. Hydrolysis of Stachyose. Upon warming with acetic acid, or even upon prolonged boiling with water, stachyose is hydrolyzed into
d-fructose
and mannatrisaccharide.
C24H4 2 O21
Stachyose
+ HÔ
= CeHÔe
d-Fructose
+ Mannatrisaccharide. ClgHs2pl6.
is
Upon warming with dilute hydrochloric or sulphuric acid stachyose rapidly hydrolyzed into its component monosaccharides. 3 HÔ = CeHÔe 2 CeHÔe. C24H42021 CeHÔe
Stachyose
d-Fructose
d-Glucose
d-Galactose.
The theoretical yield of reducing sugars from stachyose anhydride according to the preceding equation is 108.1 per cent; in actual practice, however; this yield is never reached owing to destruction of fructose.
Winterstein* obtained as a maximum, after heating stachyose 1 hour with 2 per cent hydrochloric or sulphuric acid, only 80.14 per cent yield of reducing sugar which is less than 75 per cent of the theoretical. Fermentation. Stachyose is only partially fermented by yeast;
invertase hydrolyzes the sugar into d-fructose and mannatrisaccharide, the former being quickly and the latter only slowly and imperfectly
fermented.
Lupeose. Lupeose, which was originally regarded as a galactan and afterwards as a disaccharide, is, according to the latest researches
*
Landw. Vers. Stationen,
41, 375.

of Schulze,* in all probability a tetrasaccharide, knowledge the sugar is placed in this class.
749
For want of other
but
Lupeose was discovered by Bey erf in lupine seeds preparation in a pure form was due first to Schulze { and his coworkers. The sugar occurs as a reserve substance in the seeds of
Occurrence.
its
Lupinus
luteus, L. angustifolius, etc.,

first
and
is
completely metabolized
few days of germination. Finely ground lupine seeds are extracted with 80 Preparation. per cent alcohol and the filtered extract freed of impurities by successive treatments with tannic acid, lead acetate and phosphotungstic acid. After removing the excess of clarifying agents (see under stachyose) the The precipisolution is evaporated and treated with absolute alcohol. tated lupeose is purified by dissolving in water and reprecipitating with alcohol as described under stachyose. The final product is dried
during the
over concentrated sulphuric acid.
Lupeose consists of a white, amorphous, hygrowhich has not been obtained as yet in crystalline form. scopic powder, It is easily soluble in water, less soluble in 80 per cent alcohol, insoluble in absolute alcohol and ether. Lupeose is strongly dextrorotatory. = + 148.0. of Schulze measurements latest to the [a] D According and does not of dilute alkalies solutions is not affected by boiling Lupeose
Properties.
reduce Fehling's solution.

of
mucic
is
acid.
Upon
Oxidation with nitric acid gives a large yield boiling with dilute hydrochloric or sulphuric acid
lupeose
hydrolyzed into a mixture consisting of d-galactose, d-fructose and d-glucose, the former to the extent of about 50 per cent. This would correspond to a tetrasaccharide made up of 2 molecules of d-galactose
and
is
molecule each of d-glucose and d-fructose; additional investigation required, however, before the composition of lupeose can be expressed
1
with certainty.
Verbascose.
in the roots of the
This sugar, discovered by Bourquelot and Bridel common mullein (Verbascum Thapsus), has been
||
classified provisionally as
Preparation.
hol.
The sugar is hydroxide solution; the insoluble barium compound

* f
a tetrasaccharide. Fresh mullein roots are extracted with boiling alcoprecipitated from the concentrated extract by barium
is
filtered off,
de-
Ber., 43, 2230.
Landw. Vers. Stationen, 9, 117; 14, Schulze and Steiger, Ber., 19, 827;
Ber., 43, 2233.
164.
20, 280, Schulze
and Winterstein,
Ber.,
25, 2213.
II
Compt. rend. 161,760.
750
SUGAR ANALYSIS
composed in water with carbon dioxide, and the solution of sugar The filtered; any excess of barium is removed with sulphuric acid. filtered solution is concentrated and treated with a large excess of 95 per cent alcohol which causes a precipitation of the sugar. The latter The is filtered off, and c(ried in vacuo over concentrated sulphuric acid.
sugar is purified by dissolving in hot methyl alcohol (diluted one-fifth with water), filtering and then adding one-half the volume of absolute
alcohol.
The verbascose
crystallizes
is
upon
cooling.
obtained as small needle-like crystals of sweetish taste, soluble in water, but almost insoluble in strong alcohol. The crystals, after drying in vacuo over concentrated sulphuric acid,
Properties.
Verbascose
lose 2.37 per cent of water of crystallization at 100 C. The sugar melts at 219 to 220 C. (Maquenne's Block) and at 213 C. (capillary
tube).
Verbascose
is
dextrorotatory,
[a] D (for
the sugar dried at 100 C.)
169.9, and does not reduce boiling Fehling's solution; it is only partially hydrolyzed by invertase and, upon oxidation with nitric acid, yields mucic acid equivalent to 56.7 per cent galactose; d-glucose and
= -f-
d-fructose are obtained as other products of hydrolysis. Verbascose is a true isomer of from which it differs in higher apparently stachyose
melting point and in higher specific rotation.
CHAPTER XXII
THE AMINO SUGARS AND THE CYCLOSES
IN addition to the monosaccharides, previously described, there are a number of closely related compounds which from their frequent association with the ordinary sugars and their similarity in properties have more than a theoretical interest for the analyst. Only two classes of
substances will be considered in this connection, the amino sugars and the cyclic sugars; in the description of these only such compounds will
be mentioned as
may
be met with in the investigation of plant and
animal substances.
THE AMINO SUGARS

considerable theoretical interest as they form one of the connecting links between the carbohydrates and the Only one compound, aminoglucose or d-glucosamine, will be proteids.
described.
The amino sugars have
For an account of the
many
ence should be
made
to the special
synthetic amino sugars works upon the subject.*
refer-
D-GLUCOSAMINE.
Chitosamine.
CH OH HOCH HOCH HCO H

2
A HNH
CHO
Occurrence.
d-Glucosamine does not occur in nature, so far as

is formed, however, during the hyof animal and vegetable substances nitrogenous
it
known,
drolysis
origin.
in the free condition;
of
many
the animal substances which yield glucosamine upon hythe most important are the mucins or mucoids and the chitins. drolysis The mucin of human sputum yields upon hydrolysis with hydrochloric acid about 34 per cent of the weight of dry substance as glucosamine
Among
For a
full
drates, see article
of the amino sugars and carbohydescription and bibliography Biochem. the in Handlexikon, p. 527. Geza Zemplen by
751
752
chloride;
SUGAR ANALYSIS
mucins from other products of the body also yield large quantities of the same compound. Among the mucoids the ovomucoid of eggs, the chondromucoid of cartilage and the mucoid of blood serum have been examined and these yield in some cases as high as 30 per cent glucosamine chloride. The material which yields the largest amount of glucosChitin. amine upon hydrolysis is chitin,* a nitrogenous substance found in the
members
outer covering of lobsters, crabs, scorpions, spiders, insects and other Chitin is also found widely distributed in of the Arthropoda.
the vegetable kingdom, as a constituent of the cellular tissues and membranes of the lower orders of plants, such as lichens, mushrooms, moulds,
Chitin, when purified, yields over fungi, bacteria, etc. its weight in glucosamine chloride.
80 per cent of
The
it is
also uncertain
exact chemical nature of chitin has not as yet been determined; whether the chitins of different origins are identical
in composition or are condensations of
glucosamine with varying com-
plexity.
Arakif ascribed to the chitin of lobster shells the formula
Ci8H3oN2 Oi2.
acetic acid
is
8
Upon heating this with concentrated potassium hydroxide,

split off
30
2
with formation of chitosan.t

2
Ci H N Oi2 + 2 H Chitin
CH COOH + Acetic acid

3
Ci 4
H N Oi
26
2
Chitosan.
Chitosan
it is
properties;
a yellow amorphous substance with pronounced basic upon heating with concentrated hydrochloric acid to 110 C. rapidly hydrolyzed, yielding acetic acid and glucosamine chloride.
is
CH OH
2
C 14H 26N
Oi
+ 2 HC1 + 2 H O = CH COOH + 2
2
(CHOH)s
CH- NH HC1
2
Chitosan
Acetic acid
Glucosamine chloride.
is
According to Irvine
the formula of chitin
C oH 50 Oi
3
N4, the hy-
drolysis with hydrochloric acid proceeding as follows: 4 HC1 = 4 C 6 13 5 NHCl+3 7 CaoHwOwN* 2O
+ H
CH COOH
3
Chitin
Glucosamine chloride
Acetic acid.
Glucosamine chloride is most easily Preparation of Glucosamine. prepared from lobster shells; the latter are first pulverized and then washed in cold hydrochloric acid in order to remove mineral matter. The crude chitin thus obtained may be still further purified by warming with dilute alkalies and extracting with alcohol and ether. The ex*
Discovered by Odier in 1823 (Memoire, Soc.
hist, natur.
de Paris,
1, 35).
t Z. physiol.
Chem., 20, 498.

28, 82.
J Hoppe-Seyler, Ber., 27, 3329;

J.
Chem.
Soc., 96,
564-570 (1909).

tracted material
is
753
acid until solution
is
then heated to boiling with concentrated hydrochloric effected; the liquid is then diluted, decolorized with
bone black,
filtered
and evaporated when the glucosamine chloride
will
separate as brilliant shining crystals. recrystallizing from 80 per cent alcohol.
The compound
is
purified
by
Glucosamine chloride has a sweet taste with a bitter
after-flavor.
[a]
Its solutions are strongly dextrorotatory, showing mutarotation; after solution 100 about and constant 72.5 (values
=+
[0:]^
=+
given
70 to +75). d-Glucosamine is liberated from its chloride by decomposing the latter in absolute alcohol with diethylamine, according to the method of Breuer,* or by treatment of the chloride with sodium methylate in
absolute methyl alcohol according to the method of Lobry de Bruyn and van Ekenstein.f The presence or formation of water during the process
range from
must be excluded.
Free d-glucosamine forms a fine white crystalline comC. with decomposition. It is stable in a dry atmosphere, but decomposes in presence of moisture with evolution
Properties.
pound melting
of
at about 110
forming an alkaline solution; and methyl alcohols but insoluble in ether. d-Glucosamine is dextrorotatory, [0:]^ = + 44 (Lobry de Bruyn) and + 47 to + 50 (Breuer). It is not fermented by yeast although readily attacked by moulds and bacteria. Tests. d-Glucosamine or its chloride reduces Fehling's solution and
It is easily soluble in water,
it is
ammonia.
also soluble in hot ethyl
other metallic salt solutions with great readiness, acting even in the cold. Warming with sodium hydroxide causes strong evolution of ammonia
with rapid darkening of the solution and formation of caramel-like odors. d-Glucosamine upon careful oxidation with bromine is changed to d-glucos-
COOH. aminic acid which has the formula CH 2 OH (CHOH) 3 2 Oxidation with nitric acid causes a splitting off of the 2 group with formation of isosaccharic acid. Sodium amalgam and other reducing
agents seem to have no action upon glucosamine. The ordinary color reactions of the aldose and ketose sugars also fail to develop.
CHNH NH
d-Glucosamine gives a large number of derivatives and substitution 2 group is split off and products. Heated with phenylhydrazine the in identical an osazone is formed which is every respect with that of to establish the conserves reaction This d-glucose and d-fructose.
NH
figuration of d-glucosamine. Synthesis of d-Glucosamine.
amine has been confirmed by

*
its
configuration of d-glucosFischer synthesis from d-arabinose.

t Ber., 31, 2476.
The
Ber., 31, 2193.
754
SUGAR ANALYSIS
and Leuchs* by treating d-arabinose with ammonium cyanide obtained

the following reaction:
CH OH
2
CH OH
2
HOCH HOCH HCOH CHO

d-Arabinose
+ NH CN =
4
HOCH HOCH HCOH CHNH

.
+HO
2
d-Glucosaminic acid
nitrile.
upon saponification yields d-glucosaminic acid, the lactone of which upon reduction is converted into d-glucosamine. The above reaction may serve as a general example for the synthesis
nitrile
The
of
amino sugars.
Chitose.
C Hi
6
5.
d-Glucosamine chloride, when dissolved in water Preparation. and shaken up in the cold with a slight excess of silver nitrite, loses its 2 group and by a process of inner condensation is converted into
NH
chitose.
CH OH CHOH
2
CH OH
2
CH-CHOH
HOH) 2
k HNH HC1
2
AgNO =
2
AgCl
+2H
+N
CH-CHOH
CHO
Chitose.
CHO
d-Glucosamine chloride
Chitose has been obtained only as a colorless dextronon-fermentable rotatory sirup, all attempts to crystallize it having thus far proved unsuccessful. The above constitution, proposed by
Properties.
Fischer and Andrese,f is based the analysis of its derivatives.
upon the reactions
of chitose
and upon
Chitose in
many
tion of hydrazones,
oxime reaction,
of its properties, such as reducing power, formaetc., behaves as an ordinary reduc-
On the other hand, in its failure to form osazones, chitose ing sugar. does not behave in a manner typical of the normal monosaccharides,
and
this is
supposed to be due to the absence of a
HCOH
group
in
the position adjoining the
CHO
radical.
Chitose was
acids
first
upon
chitin.
The
observed by Berthelott in the action of mineral chitose thus obtained seems to have been due,
2587.
rend., 47, 227.
however, to the decomposition of glucosamine.

*
Ber., 36, 3787; 36, 24.
f Ber., 36,
Compt.

Reactions.
755
Chitose upon oxidation with bromine
is
converted into
chitonic acid, CeHioOe, and upon oxidation with nitric acid into isosacThe configuration of these follows from that of charic acid, C6 8 7
chitose
CH OH
2
COOH
CH-CHOH X CH-CHOH
COOH
Chitonic acid
CH/-CHOH
v X
CH-CHOH
COOH
Isosaccharic acid.
48 about). The two acids do not melting at 184 to 185 C. ([a] D = form lactones and cannot be reduced by means of sodium amalgam.
Chitonic acid was obtained by Fischer and Tiemann* as a sirup = 44.5), and isosaccharic acid as a white crystalline compound (W/>
Isosaccharic acid in presence of dehydrating agents
is
converted into
dehydromucic acid and gives the characteristic reaction of this when heated with sulphuric acid and isatin (p. 781). Chitose, chitonic and isosaccharic acids can be regarded as hydrated derivates of furfuran which has the formula
t
/C=CC=CI
Their close relationship to furfural and

elsewhere
(p. 782).
its
derivatives
is
referred to
THE CYCLOSES
The cycloses f are an important group of compounds, widely distributed in nature and forming a connecting link between the sugars
and the aromatic benzol-ring derivatives. The cycloses frequently occur in nature associated with the sugars and there seems to be an intimate physiological connection between the two groups of substances;
the transformation of the one group into the other has not, however, been accomplished as yet in the laboratory. Although a number of
the cycloses are isomeric with several of the sugars, the cycloses are not sugars in the chemical sense, as they contain no aldehyde or ketone group and give none of the characteristic sugar reactions.
*
Ber., 27, 138.
full description and bibliography of the cycloses see article by Viktor Grafe in the Biochemisches Handlexikon, p. 551. t
For a
756
SUGAR ANALYSIS
The cycloses may be regarded chemically as derivatives of hexamethylene, or hexahydrobenzol, which is a cyclic carbon compound of the formula:
H
C
/
HC
2
CH
CH
HC
2
A compound answering to the properties of Betite, C 6 8 (OH) 4 a tetroxyhexamethylene was found by Lippmann* in the end products of beet molasses and was hence given the name of betite. Betite crystallizes in colorless prisms melting at 224 C. It is It has no reduceasily soluble in water and is slightly dextrorotatory.
.
ing power,
is
not attacked by boiling alkalies and upon oxidation yields
quinone.
QUERCITE.
Acorn sugar.
7
Oak
sugar.
5.
C 6 H (OH)
HO H
OH
HO
\
i\
;/ /
H
OH
C / \
HO
Pentoxyhexamethylene
Quercite, which is isomeric with the methylpentoses, C 6 Hi20 5> is widely distributed in nature, being found in acorns, cork, bark and other tissues of the oak. Of the large number of possible isomeric pentoxyis the only one at present known. Quercite was discovered by Braconnot;f it is best prepared by exThe filtered extract is tracting finely ground acorns with cold water.
hexamethylenes quercite
evaporated in vacuum at 40 C. and any sugars which are present destroyed by fermentation with yeast; the solution is then clarified by means of lead subacetate to remove tannic acid and other impurities
*
Ber., 34, 1159.
Ann. chim. phys.
[3],
27, 392.

and the
filtrate freed
757
from excess of lead by means of hydrogen sulphide.
The
clear filtered solution
upon evaporation
gives crystals of quercite
which are purified by recrystallizing from alcohol.

Quercite crystallizes in colorless monoclinic prisms C., dissolve in 8 to 10 parts of water and have a sweet taste. It is soluble in hot alcohol but insoluble in ether. Quer24.24. It is not fermented by yeast, cite is dextrorotatory, [0:]^ =
Properties.
which melt at 234
although certain bacteria are able to effect a slow decomposition. Tests. Quercite does not reduce. Fehling's solution and fails to
give any of the reactions characteristic of the sugars. Hot solutions of the alkalies are without action. Upon heating at 260 to 290 C., quercite is
decomposed into quinone,
CH
6
2,
pyrocatechin, C 6 4 (OH) 2 and pyrogallol, C 6 3 (OH) 3 which sublime with other benzol derivatives. A similar series of compounds is ob, ,
hydroquinone,
C H (OH)
6
2,
tained upon heating with concentrated hydriodic acid or fusing with
potassium hydroxide. Quercite having 5

acetates
OH
upon heating with
groups yields a corresponding number of acetic anhydride at temperatures ranging
from 100 to 150 C.
The INOSITES.
C 6 H (OH) 6
6
H H
y ^
\ / C
OH
H
Isomeric Forms.
,
C / \
OH
Hexoxyhexamethylene
The inosites, which are isomeric with the hexoses, are 6 2 6 widely distributed in both the vegetable and the animal and worlds. Of the nine possible arrangements of the groups of inosite upon the two sides of the ring plane only two of these arrange-
C Hi
OH
ments possess molecular assymetry and there would, therefore, be only two optically active isomers, corresponding to the following configurations
:
OH
OH
\OH
H/ OH
OH
758
SUGAR ANALYSIS
optically active d- and 1 inosites corresponding to the above configurations occur in nature as their methyl esters pinite and que-
The two
brachite from which they have been separated

odic acid.
by treatment with hydriis
peculiarity of the inosites
is
that none of their carbon atoms

of each
structurally asymmetric, two bonds

alike with reference to the
C atom
being connected
remainder of the ring; this apparent excepand Le Bel disappears, however, if the question is regarded from the standpoint of molecular assymmetry. This compound has not been found as yet d-Inosite, CeHÔe.
tion to the theory of van't Hoff
free in
nature;
its
methyl
is
pinite,
and
:
d-inosite
ester, however, is widely distributed as obtained directly from this by heating with con-
centrated hydriodic acid.

follows
The
reaction proceeds quantitatively as
CH
6
(OH) 5 (OCH 3 )
Finite
HI = C 6 H 6 (OH) 6
Hydriodic
acid
d-Inosite
CH,I.
Methyl
iodide,
Properties.
d-Inosite consists of small colorless octahedral crystals
which melt at 247 to 248 C., and are easily soluble in water, less soluble in alcohol, but insoluble in ether. By crystallizing from water a 2 H 2 0. hydrate has been obtained having the formula C 6 Hi 2 6 d-Inosite is dextrorotatory without mutarotation, [a] D = 65 it is not fermented by yeast, and does not reduce Fehling's solution. All of the inosites upon oxidation with nitric acid yield Tests. In carrying out this test the method colored oxyquinone derivatives. A small quantity of the material to be of Scherer* is generally used. tested is treated with a little nitric acid and evaporated upon the water bath almost to dryness a little ammoniacal barium chloride or calcium chloride solution is then added and the solution again evaporated. If inosite is present a beautiful rose red color will develop; 0.5 mg. of inosite may be detected in this way. Seidelf has modified this test by using ammoniacal strontium acetate to develop the color and in this way 0.3 mg. of inosite may be detected. d-Inosite when heated to boiling with an excess of acetic anhydride in presence of a little zinc chloride is converted into the hexacetate C6 H6 (CH 3 COO)6, which is obtained as an amorphous mass insoluble in water but soluble in alcohol ([a] D = + 9.75).
Pinite, C 6 6 (OH)5(OCH 3 ). This, the methyl ester of d-inosite, isomeric with the methylhexose sugars and is found widely distributed in nature. It was discovered by BerthelotJ in 1856 in the
is *
Ann., 73, 322; 81, 375.

|
Chem.
Ztg., 11, 676.
Compt.
rend., 41, 392.

resin of the
in
759
Pinus lambertiana of California;
it
also occurs as sennite*
Senna
leaves, as matezitef in the juice of the
plant (Mateza roritina)
and has
also
Madagascar rubber been found in the mother liquors t
from the crystallization of coniferin. The identity of these various methyl esters of d-inosite with pinite has been established by Combes, Wiley, and others.^ Pinite forms white rhombic-hemihedral crystals melting at 185 to 186 C., and subliming without decomposition at 200 C. It has the same degree of sweetness as cane sugar, is easily soluble in water, less soluble in alcohol and insoluble in ether. It is not fermentable, and
||
Pinite is dextrorotatory, [a] D = +65.5. This 1-Inosite, compound has been found as yet of in the form its only methyl ester, quebrachite, from which it was obtained by Tanret ** upon heating with hydriodic acid. The reaction
does not reduce Fehling solution.
Hi20 6
is
the same as that given for pinite.

Properties.
2
from alcohol as the anhydride 247 C. A hydrate, has been obtained by crystallizing from water. 2 6 6 + 2 H 2 O, 1-Inosite is easily soluble in water, less soluble in alcohol but insoluble in ether. It is levorotatory, [a] D = 65 for the anhydride without muis unfermentable and does not reduce Fehling's solution. tarotation,
1-Inosite crystallizes
C Hi O6 C Hi
6
in the
form of
colorless prisms melting at
1-Inosite gives Scherer's inosite reaction
upon heating with
nitric acid.
With
acetic anhydride
pound is same as the hexacetate

Quebrachite,
an amorphous hexacetate is formed; the com10) but in other respects behaves the levorotatory ([a] D =
of d-inosite.
6
(OH)5(OCH 3 ). This, the methyl ester of occurs in the bark of the Quebracho tree. It crystallizes in prisms melting at 186 to 187 C.; the crystals are very sweet, easily soluble in water, less soluble in alcohol and insoluble in ether. Quebra6
1-inosite,
= 80; this figure, though of opposite sign, is levorotatory, [a] D not of the same value as that of pinite (+ 65.5), so that the two compounds are not optical antipodes. The compound is not attacked by dilute alkalies or acids; heated with concentrated nitric acid it gives Scherer's reaction. Quebrachite is unfermentable and does not reduce
chite
is
Fehling's solution.
*
t
Dragendorff and Kubly, Ztschr.
f.
Chemie
(1866), 411.
II
Compt. rend., 77, 995; Tiemann and Haarmann, Ber., Compt. rend., 110, 46. Amer. Chem. Jour., 13, 228.
Girard,
110, 84. 7, 609.
" f See Maquenne's Les Sucres," ** Compt. rend., 109, 908.
p. 209.
760
d, 1-Inosite. *
SUGAR ANALYSIS
Racemic inosite was obtained by Maquenne and by dissolving and crystallizing equal parts of d- and 1-inosite.
consists of colorless crystals melting at 253
Tanret
The anhydride
ture,
C.
the subreacts
stance behaves as a true racemic combination

d, 1-Inosite is optically inactive
;
and not
as a simple mixit
;
in its chemical behavior
not fermented by yeast it has the same Tanret who found that Aspergillus niger resolved been partially by at low temperatures caused the inactive solution to become sensibly
as either d- or 1-inosite.
It is
levorotatory.
Occurrence. i-Inosite, CeHÔe (Phaseomannite, Nucite, Dambose) Inactive inosite, also called anti- or mesoinosite, is the only inosite which has thus far been found free in nature. It was discovered by
.
Schererf in 1850 in the mother liquors from a preparation of creatine obtained by extracting meat, and has since been found to be very widely distributed throughout the animal and vegetable kingdoms. It
occurs in the muscles, kidneys, liver, lungs, heart, brain and other organs of the body and has also been found in the urine of patients
afflicted with Bright's disease and diabetes, and also frequently in normal urines. The occurrence of inosite in the urine is sometimes termed inosuria. In the vegetable world i-inosite has been found in green beans, peas and other legumes, in the cabbage, in the leaves of asparagus, the potato, dandelion, grape vine, oak, ash and other trees, in different mushrooms, in the roots of many plants and in the juices of grapes, blueberries and other fruits. In the combined form i-inosite occurs as its methyl esters bornesite and dambonite. i-Inosite is prepared from meat by first extracting Preparation. the finely cut material with water. The aqueous extract is then slightly acidified with acetic acid and boiled; the coagulated albumin is filtered off and the filtrate clarified with normal lead acetate. The solution is again filtered and the filtrate heated with an excess of lead subacetate solution and allowed to stand for 1 to 2 days. The basic lead-inosite compound is filtered off and decomposed in water with hydrogen sulphide. The filtrate from the lead sulphide is concentrated, treated with an excess of hot alcohol and the solution separated from
The alcoholic solution upon cooling will usually deposit crystals of inosite; if no crystals form, the separation may be promoted by adding ether to the point of turbidity, and setting
any precipitated impurities.
* t
Compt.
rend., 110, 86,
Ann., 73, 322.

the solution aside in a cool place.
761
The compound
is
purified
by
re-
crystallizing from alcohol. To prepare inosite from plant materials the process employed by Maquenne* for its extraction from walnut leaves may be employed. The dried finely ground leaves are extracted repeatedly with 5 to 6 parts of boiling water, the residue pressed out and the brownish colored extract treated hot with concentrated milk of lime until the precipitate which has formed settles quickly. The solution is filtered and the filtrate treated with a very slight excess of normal lead acetate. The solution is again filtered and the inosite precipitated with ammoniacal The precipitate, which should be perfectly lead subacetate solution. then off and filtered is decomposed in aqueous suspension with white, from the lead sulphide precipitate is filtrate hydrogen sulphide. The then treated while still warm (about latter is a the to sirup; evaporated 50 C.) with 7 to 8 per cent of its volume of concentrated nitric acid which oxidizes most of the impurities but is without action upon the inosite. (Excess of acid and high temperature must, however, be The acid solution is then heated for a few minutes upon avoided.) the water bath and then treated with 4 to 5 volumes of strong alcohol; after cooling 1 volume of ether is added when the inosite will begin to After 24 hours the solution is decanted, the impure inocrystallize. site washed with alcohol and then recrystallized from acetic acid. To remove the last traces of coloring matter, calcium sulphate and other impurities, the inosite is dissolved in water and treated with a slight The solution is filtered, the exexcess of barium hydroxide solution. cess of barium removed with ammonium carbonate and the clear filtrate evaporated to dryness. The residue upon recrystallizing from water gives pure inosite. By this method Maquenne obtained 440 gms. of inosite from 150 kgs. of leaves, a yield of about 0.29 per cent. i-Inosite crystallizes from alcohol or from water above Properties. a temperature of 50 C. as the anhydride in the form of needles melting at 224 C. Upon crystallizing from water below a temperature of 50 C., the hydrate CeHÔe + 2 H2 is obtained in the form of large hexagonal monoclinic crystals which effloresce rapidly in a dry atmos-
phere. at 15
ether.
i-Inosite has a sweet taste, is very soluble in water (7.5 parts C. for the anhydride), less soluble in alcohol and insoluble in It is optically inactive even after the addition of borax; its
is
not affected by the attack of moulds as is the case is not fermented by yeast, although certain bacIt does not reduce Fehling's teria appear to cause destructive changes.
optical neutrality
with
d, 1-inosite.
It
* "
Les Sucres,"
p. 216.
762
reagent, although
solution.
it
SUGAR ANALYSIS
produces a metallic mirror with ammoniacal silver
i-Inosite gives Scherer's reaction, described
under
d-inosite.
Bornesite.
i-inosite,
Borneo caoutchouc; it in the wash waters from certain was also rubber factories. It is isomeric with pinite and quebrachite and crystallizes in rhombic prisms melting at about 200 C. and subliming at 205 C. It is easily soluble in water, but less soluble in alcohol. It is = + 32 (Girard), + 31.16 (Flint and Tollens); it is dextrorotatory, [a] D unfermentable and does not reduce Fehling's solution. It is decomposed J by heating with hydriodic acid into methyl iodide and
in crude
i-inosite.
(OH) 5 (OCH 3 ). was discovered by Girard* found by Flint and Tollensf

6 6
This, the
monomethyl
ester of
This, the dimethyl ester of Dambonite, CeHetOHWOCI^. was discovered by Girard in Gabon rubber; it has also been found in the latex or milky caoutchouc yielding juice of the Castilloa Dambonite crystallizes in white rhombic prisms which melt elastica. at about 190 to 195 C. and sublime between 200 to 210 C. It is sweet, very soluble in water and dilute alcohol, unfermentable, optically inactive and does not reduce Fehling's solution. Dambonite forms with potassium iodide a double salt of the formula C 8 Hi 6 6 KI. Upon heating with hydriodic acid it yields methyl iodide and i-inosite. Hyi-inosite,
drolysis
acid.
is
also effected
upon heating with concentrated hydrochloric

.
This compound was discovered by VinQuercinite, C 6 6 (OH) 6 cent and Delachanal in the mother liquors obtained from the crystallization of quercite. Quercinite crystallizes from cold water as a
||
hydrate, the crystals of which effloresce rapidly upon exposure to the air. Crystallized from hot water the anhydride is obtained in the
form of rhombic prisms melting at 340 C.

in
The anhydride is soluble 66 parts of cold water, easily soluble in hot water, insoluble in alcohol and ether; it is optically inactive, unfermentable and does not reduce
Quercinite gives Scherer's inosite reaction, and in general behavior seems to belong to the group of inactive inosites of which there are seven possible stereo-isomers.
Fehling's solution.
its
Phytin.
Inosite also exists in nature in combination with phos-
phoric acid as phytin, the principal phosphorus

*
compound
of vegetable
Compt.
rend., 73, 426; 77, 995.

12, 566.
t Ann., 272, 288. J
II
Maquenne, Ann. chim. phys. Compt. rend., 67, 820. Compt. rend., 104, 1855.
[6],

seeds.
763
Phytin, according to the researches of Suzuki, Yoshimura and
Takaishi,* is an inosite-hexaphosphoric acid C 6 6 [OPO(OH) 2 ]6 which, by the action of a special enzyme phytase, is hydrolyzed into inosite and
,
phosphoric acid.
C6H [OPO(OH)
6
2]
= C
Hi 2
H P0
3
4.
Phytin
Inoaite
Phosphoric
acid
Bull. College of Agric.,
Tokyo,
7, 495,
503 (1907),
CHAPTER XXIII
THE SUGAR ALCOHOLS AND SUGAR ACIDS
THE close relationship of the sugars to the alcohols upon the one side and to the monobasic and dibasic acids upon the other has already been mentioned. While these two groups of substances are entirely distinct from the sugars, their constant association with the sugars in nature and their great importance in many analytical and synthetical operations of sugar chemistry are of sufficient account to require brief
mention.
THE SUGAR ALCOHOLS

Of some thirty known sugar alcohols the following eight have been found in nature: glycerol, erythrite, adonite, sorbite, mannite, dulcite, Reference has already been made to the perseite and volemite.
occurrence of these. The sugar alcohols are generSynthesis of the Sugar Alcohols. the of nascent action hydrogen upon an aldose or ally prepared by
ketose sugar.
amalgam.
The reduction is best accomplished by means of sodium The process of Fischer* is as follows: a 10 per cent aqueous
solution of the sugar is treated ice cold with small additions of sodium amalgam (2 to 2| per cent sodium content) until the reducing power of the solution has almost disappeared. During the first part of the
operation the solution is kept weakly acid with constant additions of dilute sulphuric acid in order to prevent molecular transformation of
sugar by action of the free alkali; in the last stages of the reduction the solution is kept faintly alkaline. After reduction the solution is
neutralized, evaporated until sodium sulphate begins to crystallize and then poured into 8 volumes of absolute alcohol. The alcoholic solu-
tion
alcohol
from sodium sulphate and evaporated when the sugar obtained either as a sirup or in crystalline form. The sugar Formation of Sugar Alcohols During Fermentation. alcohols are also formed in many anaerobic fermentations through a
is filtered is
The best-known example of this is the similar process of reduction. so-called mannitic fermentation which takes place frequently in the
juices of the sugar cane, sugar beet, grapes, apples and in other vege" * " (1909), pp. 186, 292, 473, etc. Untersuchungen uber Kohlenhydrate 764
765
The sugar is changed partly to mannite and partly to table extracts. the mucilaginous gum dextran (C 6 Hi O 5 ) n the latter can be precipitated by means of alcohol and the mannite obtained by evaporation of
;
the alcoholic solution.
The presence
gars, sugar-house products, distillery
mannite in wines, musts, vineresidues, etc., is due largely to the

of
result of such fermentations.
The sugar alcoProperties and Reactions of the Sugar Alcohols. hols resemble one another in their sweet taste, in not being fermented by yeast and in the complete lack of the aldehyde or ketone properties
(reduction of Fehling's solution, hydrazone and osazone formation, color reactions, etc.), characteristic of the parent sugar. In presence of free alkalies the sugar alcohols give soluble complex substitution products with
many of the heavy metals;
for this reason salts of copper, etc.,
by alkaline hydroxides in presence of glycerol, mannite and other polyvalent alcohols. This property, however, is not a characteristic one, being also shared by the sugars and their acid
derivatives.
If excess of alkali be of Sugar Alcohols and Metals. metallic substitution the of the products avoided, sugar alcohols may be obtained in some cases as a precipitate. Mannite, for example, can
are not precipitated
Compounds
be precipitated from solution in presence of copper sulphate by adding hydroxide to faintest possible excess. The blue coppermannite compound can then be filtered off; it is practically insoluble in
ammonium
water, but
is soluble in excess of ammonia from which solution the mannite can be regenerated after removing the copper with hydrogen * sulphide. This process due to Guignet can be utilized for the separation of mannite from plant juices.
The behavior of many Reaction of Sugar Alcohols with Borax. borax and boric acid is with also alcohols worthy of mention. If sugar
a
little
borax be added to an aqueous solution of mannite, arabite,
etc.,
the solution becomes strongly acid, with a

trical
marked
increase in the elec-
The phenomenon is due to the formation of conductivity.! alcohol-boric acid complexes, the constitution of which remains in The
acid complex, which
is
doubt.
strong enough to decompose car-
.bonates, undergoes dissociation J upon dilution with water. Borax and boric acid also have the peculiar property of intensifying
the rotatory power

*
of solutions of the sugar alcohols to a very

rend., 109, 528.
Ital.,
marked
Compt.
Magnanini, Gazetta chim.

chim.
[2],
20, 428.
} Klein, Bull. soc.
Vignon, Compt.
29, 195, 198, 357. rend., 77, 1191; 78, 148.
766
degree, the result alcohol complex.
SUGAR ANALYSIS
no doubt
of the higher specific rotation of the boric acid
Acid molybdates * of sodium and ammonium produce the same effect to an even greater extent; so also the tungstatef and paratungstate of sodium. Polarization of solutions before and after the addition of constant quantities of borax has been employed for estimating certain sugar alcohols, as mannite, i in mixture with other substances.
Table CIII gives a list of the different alcohols, with a few of their properties, which are obtained by reduction of the different monoThe sugar alcohols, which have been found free in saccharides.
In the nomenclature of the sugar nature, are marked in italics. alcohols the ending -ite is usually substituted in place of the termination -ose of the sugar, as pentite, hexite, etc. It will be noted from the table that the ketose sugars, erythrulose,
fructose, sorbose,
tion.
and tagatose, yield two isomeric alcohols upon reduc-
This necessarily follows from the configuration, since reduction of

will give
the
C = O group C=O
HCOH and both HC(
HOCH
isomers.
A number of reReaction of Sugar Alcohols with Aldehydes. which have been employed for the separation and identification Chief among these are the of the sugar alcohols, should be mentioned. reactions with formaldehyde, acetaldehyde and benzaldehyde in presence of strong hydrochloric or sulphuric acid (50 per cent) with formaactions,
tion of a characteristic group of
compounds known as acetals. Formats. Mannite, for example, when heated with equal parts of 40 per cent formaldehyde and concentrated hydrochloric acid gives
mannite triformal,
slightly soluble in
||
C6H8
(CH 2 )3, which
consists of white needles, only
C. water and In the same way, by heating mannite with acetaldehyde or paracetaldehyde in presence of concentrated hydrochloric acid or 50 per cent sulphuric acid, mannite triacetal, CeHsOe^Hâ, is formed.
melting at 227
Acetals.
Benzols.
tion
*
Of greater value than the formals and acetals
for separa-
and
identification of the sugar alcohols are the benzals.
This re-
Gernez, Compt. rend., 112, 1360.
t Klein,
Compt.
rend., 89, 484.
j Muller, Bull. soc.
chim.
[3],
11, 329.
chemists prefer the ending -itol in place of -ite, as mannitol, arabitol, perseitol, etc.; while this conforms with the rule that all alcohols should end in -ol the author has preferred the older and simpler terminology, which is still retained by
Fischer, Tollens,
II
Many
Lippmann, Maquenne and other leading
authorities,
Tollens and Schulz, Ber., 27, 1892.
767
ffff
oo
II
ill Iff *
00
+ +
W
OOO
8
8
||
OO
"^ "n
_rt
OOO
"o
1 s
lO iO 10 t^ rH
i
i
2
'
-
oj
'5*!a
II 1 o<
O
-
1 1 1 Mtn'S
(DQJO)
-
II
O>O>
-
HH
S 6 1 1 qs1d1 11~q ddddd d d dddod dd dddo
q qqqqq d ddddd
a
wwwww
.-
|llfl
lll,
111
-
(o
gg
ill if
lf
ii j=ii
1 1
T?i^4
&
TS-OrtPn
$Sj
I "^
I
"T3
Illll T?
'i
T?
'i
Tl
768
SUGAR ANALYSIS
S
1
ro
H>
pecific
CO
+000 +1+
t
S &
CO
O CO
++53
++++
o
t**
OO OO to
o
T-H
o
1-H
<*<
OO Ô 10 <M O5
02
-a
-+J
& o
42
^2o2
"a >>
S
.1 43
a JnO)
os cjo
434343
qqq WWW u o
c5
-a
'W qqqqqqqqqgq q qqq WWWWWWWWW^W W WWW
qqq WWW
'
*3
-^>
fe
ft,
^ 2
C=3o
J-3^ Soo
w o q O W cT
r ? '
< fl
PP
U7
r^
2s" W o r? Q U

action,
769
which
is
due to Meunier,* has been much employed by Fischer. f
The
acid,
alcohol, in concentrated hydrochloric acid or 50 per cent sulphuric

is
shaken up with benzaldehyde when the benzal derivatives of and perseite will quickly precipitate: the separation with these alcohols is almost quantitative. In the case
erythrite, xylite, mannite, sorbite,
of glycerol, arabite, and dulcite the benzal derivatives obtained by this method remain in solution so that no separation is effected. As to the constitution of the benzals obtained by the method just described there appears to be no uniformity. Mannite, for example,
combines with three molecules of benzaldehyde; erythrite, xylite, adoand perseite with two; and glucoheptite with only one. This peculiarity is probably due to the spatial arrangement of the alcohol groups within the molecule, although no satisfactory theory J has as yet been formulated. As in the case of the formals and acetals the reaction probably results from the withdrawal of the from 2
nite, sorbite,
hydroxyl groups of the sugar alcohol by the reaction, for example, with sorbite would be:
of the aldehyde.
\
The
CeHÔo
Sorbite
H 2 O C-H = C H O
C
6 5
:
CH
6
Benzaldehyde
6 10 6\ : Sorbite-dibenzal
C -H/
H O, +2 Water.
2
reaction
but which of the hydroxyl groups of the sugar alcohol participate in the is not at present known.
more important benzal derivaTable CIV. The benzals upon boiling with 5 per cent sulphuric acid are decomposed into benzaldehyde and the free alcohol. The process of decomposition is much facilitated by the addition of a little free benzaldehyde. In a few cases, as with mannite, long boiling and a high temperature of
properties of the
tives of the sugar alcohols are given in
The formulae and
heating are required to effect complete hydrolysis. The benzaldehyde can be removed by shaking out the cold acid solution with ether and the
sulphuric acid eliminated by neutralizing with barium hydroxide and The clear filtrate upon evaporation filtering off the barium sulphate.
will
then yield the sugar alcohol either as a sirup or in the crystalline
form.
By this means it is possible to effect the separation of different sugar alcohols from plant extracts, juices, etc. Sugar alcohols can be detected in the presence of sugars by first heating the solution with dilute hydrochloric acid to invert any higher
saccharides
as osazones
*
;
by means
t
the sugars are then precipitated in the neutralized solution After filtering off the osaof phenylhydrazine.
Compt.
t Ber., 27, 1524. rend., 106, 1425, 1732; 107, 910; 108, 408. See Fischer's discussion upon this point, Ber., 27, 1524.
770
zones, the filtrate
is
SUGAR ANALYSIS
shaken out with ether to remove excess of phenylfor sugar alcohols with
TABLE CIV
Giving Formulae and Properties of Sugar Alcohol Benzols
hydrazine and the aqueous solution tested benzaldehyde in the manner described.
Alcohol.

parts nitric acid of 1.18 sp. gr. at a temperature of about 45
liquid soon acquires reducing properties
771
C.
The
and when this has reached its maximum the solution is cooled, neutralized, and the sugar precipitated as hydrazone, osazone, or examined by other suitable methods. In
place of nitric acid other agents may be used for oxidizing the sugar alcohols to sugars, such, for example, as sodium hypobromite, weak permanganate, hydrogen peroxide in presence of ferrous sulphate, and lead
peroxide with hydrochloric acid. Oxidation of Sugar Alcohols by

of the sugar alcohols to sugars
Means
of Bacteria.
The oxidation
be accomplished by biochemical means. The organism most used for this purpose is the Bacterium xylinum, or sorbose bacterium, the action of which upon sugar alcohols and sugars has been especially studied by Bertrand.* The peculiarity of the oxidation of sugar alcohols by Bacterium xylinum is that the sugars formed are largely if not entirely ketoses. The following ex-
may also
amples are given of oxidation of sugar alcohols made by means of organism:

i-Erythrite 1-Arabite
this
= = = d-Mannite = d-Sorbite = Perseite = Volemite =

Glycerol
Dioxyacetone.
Erythrulose. Araboketose.
d-Fructose.
d-Sorbose.
Heptoketose. Heptoketose.
xylinum.
All the sugar alcohols, however, are not oxidized by Dulcite and xylite, for example, are not affected
Bacterium
by
this or-
ganism. A curious fact noted in this connection is that oxidation by Bacterium xylinum does not take place in compounds where the hydroxyl groups in the second and third position lie on opposite sides of the carbon chain. Thus xylite and duleite both have the following
configuration in
common:
HO-C-H H-C-OH
CH OH
2
2
1
For some reason not understood sugar alcohols having the above
arrangement, are not oxidized by Bacterium xylinum. Sorbite, mannite, arabite and erythrite, on the other hand, have the
*
Compt.
rend., 126, 762, 894, 984; 130, 1330; Bull. soc. chim.
[3],
16, 627; 19,
347.
772
SUGAR ANALYSIS
hydroxyl groups in the second and third position on the same side of the carbon chain and are oxidized by Bacterium xylinum as follows:
3 2
1
H-C-OH H-C-OH+O
CH OH
2
H-C-OH
=
C:O
+ H O.
2
CH OH
2
Alcohol
Ketose.
Too prolonged
of
or too violent oxidation of the sugar alcohols will
lead beyond the sugars to the formation of sugar acids, the description
which
will follow.
THE SUGAR ACIDS

The sugar
two
acids according to the degree of oxidation are divided into the monobasic and the dibasic acids. groups:
THE MONOBASIC ACIDS OF THE SUGARS
The oxidation of sugars to Synthesis of the Monobasic Acids. the monobasic acids is usually accomplished by means of bromine water.
The
general equation for the reaction with an aldose sugar
is
:
C n H2n O n
Aldose
C n H O n+1 + +Bromine + H O = Aldonic

2 Br
2
2n acid
2 HBr.
Hydrobromic
acid.
1 part of sugar In carrying out the reaction according to Fischer dissolved in 5 parts of water and 2 parts of bromine added. The solution is kept cold and shaken frequently until all bromine has disis
After standing at room temperature 1 to 3 days, the solution is heated to expel any excess of bromine; carbonate of lead is then added to neutralize the hydrobromic acid formed in the reaction and the filtered
solved.
solution evaporated to about half its volume.
After 24 hours the lead
bromide, which has
crystallized,
is
filtered off
and the lead remaining
in
solution precipitated with hydrogen sulphide. After boiling off the from the filtrate the last traces of hydrobromic acid hydrogen sulphide
are
dissolved silver
removed from the solution by shaking with silver oxide, and any removed from the filtrate with hydrogen sulphide. The
solution is then reboiled to expel hydrogen sulphide, decolorized if necessary with animal charcoal and filtered when the acid can either be precipitated, in the form of an insoluble salt or other derivative, or
separated as a crystalline lactone by evaporation. The method just described for oxidizing sugars to their monobasic acids holds true, however, only for the aldose sugars. Ketose sugars are
but
little
affected
by bromine water at ordinary temperature during the

*
Ber., 22, 3218.

first
773
few days. Several weeks' contact, however, will bring about slow oxidation, with a breaking up of the molecule into a mixture of acids of lower carbon content. /Lusfrr f 4| In the nomenclature of the monobasic acids deNomenclature. rived from the sugars the ending -onic is usually substituted for the
,
termination -ose of the sugar as xylonic, gluconic, pentonic, hexonic,

etc.
Lactones of the Monobasic Acids.

collic acid,
corresponding to a diose sugar,

C.)
Of the monobasic acids glyis obtained in the form of

acid, corresponding to a
crystals (melting point 80
and gly eerie
triose sugar, as a thick sirup.
The
heptonic, octonic
and nononic acids
split off
higher tetronic, pentonic, hexonic, one molecule of water upon
evaporation and crystallize out as lactones. The formation of the lactone of a hexonic acid is represented as follows:
CH OH CHOH
2
7
ft
CHOjH CHOH
+H
CHOH
!
OC-;OH
Hexonic acid
Hexonic acid lactone
In the above reaction the splitting off of water and the linkage by the oxygen ring always take place between the carbon atom of the terminal COOH group and the third or 7 carbon atom. Glycollic and glyceric acids, which have no 7 carbon atom, are unable to form lactones. The lactones of the monobasic acids are well-defined crystalline
compounds easily soluble in water. Freshly prepared aqueous solutions of the lactones 'are neutral in reaction, but on standing a strong
acidity develops owing to the regeneration of the carboxyl group addition of water.
by
The sugar monobasic acids and their lactones are optically active. marked difference, however, is noticeable between the specific rotations of the acid and its lactone, and with the transformation of the
one into the other, by the addition or splitting off of water, changes in This rotation take place which resemble the mutarotation of sugars. is seen from the following observations made upon galactonic acid and its lactone; to reduce the influence of lactone formation the observations for the free acid were made upon a solution prepared by decomposing a
774
SUGAR ANALYSIS
amount
of oxalic acid
solution of the calcium salt with the equivalent
and
filtering.
Galactonic acid
from calcium
salt
10 minutes after solution 5 hours after solution 6 days after solution 15 days after solution 3 weeks after solution
Gfllartonir acid t lactone Lralactomc arid T lartone
I s
immediately after solution severa j days after solution
- 10.56 - 13.77 - 39.24 - 45.90 - 46.82 - 77.61 - 67.89
The observations show a slow conversion of the acid into the lactone and a similar conversion of the lactone into the acid; after a longer or shorter period of time, depending upon temperature and concentration,
a condition of equilibrium is reached when the rotation remains constant. Relation of Configuration to the Rotation of Lactones. An important relation, noted by Hudson, f between the configuration and rotation of the lactones of the sugar acids, is that all dextrorotatory lactones have their ring linkages upon one side of the carbon chain and
levorotatory lactones upon the opposite side. In the following table the position of the lactone ring, with reference to the terminal group and the carbon chain, is indicated at the head of the two
all
CO
classes of lactones.
Dextrorotatory lactones.

etc., (see
775
different saccharinic acids, as saccharin, isosaccharin, metasaccharin,
pp. 587 and 604). The relationship is an important one, since the rotation of a lactone indicates the position of the ring, thus establishing the configuration for the y position of the acid and hence also
for the corresponding sugar.
In this manner Hudson has not only
by Fischer from but has out the chemical data, pointed probable structure of a purely number of sugars whose configurations have been in doubt. A peculiarity of Molecular Rearrangement of the Sugar Acids.
verified the configurations of the sugars established
many
The
sugar acids
is
into other isomers
when
the ease with which they undergo molecular change their solutions are heated at high temperature.
is
simplest instance of such a change
the transformation of dextro-
lactic into levo-lactic acid or vice versa, the reaction as in all
such cases
being a reversible one.
CH
CH
H-C-OH^ HO-C-H HO-C=O HO-C=O

d- Lactic acid
1-Lactic acid
lactones do not appear susceptible to this kind of molecular rearrangement and to prevent their formation the experiment with
acids,
The
by heating the aqueous soluC. in presence of pyridine or quinoline, the latter through formation of salts preventing the generation of lactones. The part of the molecule which is affected in this method of isois
which yield lactones,

to 150
carried out *
tion at 130
is always the hydroxyl adjoining the carboxyl group, the for the reaction being: formula general
merization
H-C-OH _> HO-C-H 0=C-OH 0=C-OH

The following examples are given of monobasic acids which have been found to undergo mutual isomerization by the method of Fischer
just described.
1-Xylonic 1-Arabonic
<=*
d-Lyxonic
1-Ribonic
<
d-Gluconic
1-Gluconic
d-Mannonic
1-Mannonic
d-Talonic
1-Idonic
+
<=
<=
d-Galactonic
1-Gulonic
The same reaction is also obtained between the the heptonic, octonic and nononic acids.
*
a and
isomers of
776
SUGAR ANALYSIS
It has not been found possible Reduction of Lactories to Sugars. to reduce the monobasic acids in aqueous solution; the lactones* however, are easily reduced in aqueous solution by means of sodium amalgam
to sugars and after prolonged reduction to sugar alcohols. reaction for a hexonic lactone would be:
first
The
CH OH
2
CHOH HOH
:H
CH OH CHOH
2
CH OH
2
CHOH CHOH
-io
Hexonic lactone
nH
I
or
HOH CHOH CHOH

Hexoset
CHOH CHOH CHOH CHOH CHO
by treating an icereaction, according to Fischer, J is cold solution of the lactone in 10 parts of water with sodium -amalgam
carried out
(2J per cent sodium), the mixture being always kept weakly acid with The reaction is stopped when the reducing power upon sulphuric acid.
The
Fehling's solution has reached
its
The solution is then neutralized,
maximum (usually 30 to 40 minutes). decolorized with bone black and evapo-
rated to crystallization of sodium sulphate, when it is poured into 20 times its volume of hot alcohol. After cooling, the alcoholic solution is
from sodium sulphate and evaporated to a sirup from which the sugar may be separated as hydrazone or other compound according to conditions. The yield of sugar is 40 to 60 per cent of the pure lactone. The transEmployment of Method in the Synthesis of New Sugars. formation of the monobasic acids of known sugars into new isomers and the reduction of the lactones of the new acid by the process just described have been used by Fischer with great success in the synthesis of
filtered
many new sugars. The following is given as an illustration of the method

Sugar,
Monobasic acid (produced by oxidizing

In the same manner
1-Arabinose
:
777
= 1-arabonic acid Rhamnose = rhamnonic acid d-Galactose = d-galactonic acid = d-gulonic acid d-Gulose a-Heptose = a-heptonic acid = a-octonic acid a-Octose
1-ribonic acid
= isorhamnonic acid = d-talonic acid = d-idonic acid = /8-heptonic acid = /3-octonic acid
= 1-ribose. = isorhamnose = d-talose. = d-idose. = -heptose. = /3-octose.
Hydrazide* Reaction of the Monobasic Acids. Among the most important derivatives of the sugar acids, for purposes of identification and separation, are the phenylhydrazides. All of the acids derived from the sugars react with phenylhydrazine; the resulting product, however, is entirely different in chemical properties from the hydrazones
and osazones
of the sugars, resembling more the acid amides. The reaction of a hexonic acid with phenylhydrazine is given as illustration:
CH OH
2
CH OH
2
H H H H O:C-jOH + H;-N-N-C H = O rC-N-N-CgHs + H O

-.
I
(CHOH) 4
(CHOH)
Hexonic Acid
Phenylhydrazine
Hexonic phenylhydrazide.
The
reaction
is
carried out
by heating a
solution containing 1 part
of the acid in 10 parts of water with 1 part of phenylhydrazine and 1 part of 50 per cent acetic acid for three-quarters of an hour upon the water
cooled, the precipitate of phenylhydrazide filtered cold water and recrystallized from hot water off, animal a black. The hydrazides thus obtained are colorless little using the melting points of which will serve in many crystalline compounds,
bath.
The
solution
is
washed with a
little
upon heating with alkaline acid and free phenylhydraof a salt of the with formation hydroxides, Barium hydroxide is generally used for this purpose: 1 part of zine. hydrazide is treated with 30 parts of hot 10 per cent barium hydroxide The free phenylhydrasolution, boiled one-half hour and then cooled. zine is then extracted with ether, the barium precipitated with the exact
cases for purpose of identification. The phenylhydrazides are decomposed
amount of sulphuric acid and the solution filtered; the filtrate upon evaporation will yield the lactone of the acid. The monobasic acid derivatives Salts of the Monobasic Acids. of the sugars give a large number of salts with different metals, some Mention has of which have been used for purposes of identification.
been made of a few of these, in so far as they pertain to the identification of sugars, under the reactions of the individual sugars.
* Fischer
and Passmore,
Ber., 22, 2728.
778
SUGAR ANALYSIS
salts of calcium,
barium, cadmium, and lead have been employed in some cases for isolating certain of the acids. The cadmium
The
and lead
salts (the latter usually
amorphous
flocculent precipitates) are
decomposed after separation with hydrogen sulphide and the calcium and barium salts with the equivalent amounts of oxalic or sulphuric acid; the precipitates are filtered off and the liberated acid is obtained
by concentrating the filtrate. A number of the monobasic acids give
characteristic salts with
different alkaloids, as strychnine, brucine, morphine, and the various cinchona bases. The utilization of these salts in analyzing racemic
mixtures of sugar acids will be described later (p. 786). Oxidation of Monobasic Acids of the Sugars.
The monobasic
acid derivatives of the unsubstituted aldose sugars are converted by oxidizing agents (as nitric acid, 1.2 sp. gr.) into the corresponding dibasic
acids, rhamnonic, fuconic, rhodeonic, methylhexonic, etc., yield dibasic acids of one less carbon atom with loss of the methyl group.
acids; the substituted
monobasic
THE DIBASIC ACIDS OF THE SUGARS

Formation.
The
oxidation of sugars to their dibasic acids
is
usually performed by warming the sugar with 30 per cent nitric acid. The reaction only holds for normal unsubstituted aldose sugars, the ketoses being all degraded into lower oxidation products, of which
is usually formed in largest amount. The oxidation of an aldohexose sugar to its dibasic acid by means of nitric acid proceeds
oxalic acid
as follows:
CH OH
2
O:C-OH
(CHOH) 4
+ 2 HNO =
3
(CHOH) 4
-f 2
H O + 2 NO
2
H-C:O
O:C-OH
Nomenclature. The nomenclature of the dibasic acids is irregular. In some cases where there is a genetic relationship, as between the
sugars glucose, mannose,
and
idose,
and
their dibasic acids saccharic,
mannosaccharic, and idosaccharic, a certain uniformity exists; so also between the sugars galactose and talose, and their dibasic acids mucic
The family to which each acid belongs is usually inby the name of the saturated dibasic fatty acid having the same number of carbon atoms, as: malonic (3 C atoms), succinic (4),
dicated
glutaric (5), adipic (6), pimelic (7), suberic (8)
and talomucic.
and
azelaic (9).

Sugar.
779
780
SUGAR ANALYSIS
in water.
The lactone acids are nearly all crystalline compounds, easily soluble The solution of a lactone acid, neutralized in the cold with
sodium hydroxide, quickly becomes acid again through reconversion of Stable compounds of the lactone the lactone into the free acid group.
acids are for this reason
unknown.
Solutions of the lactone acids in water undergo spontaneously a partial change into the dibasic acid with establishment of a condition of
equilibrium, the predominance of lactone acid, or of dibasic acid, dependWith this transformation ing upon the temperature and concentration.
acid and
changes are noted in the rotation of the solution. In the case of saccharic its lactone acid, the following specific rotations were noted.
Saccharic acid,* after solution Saccharic acid, constant (29 days)

Saccharic acid monolactone, after solution Saccharic acid monolactone, constant (56 days)
WD.
9.1
-j-
22.7
+ 37.9 + 22.5
its
The
lactone
results
is
show that the change between saccharic acid and
a reversible one, the same condition of equilibrium being reached whichever compound is first dissolved. The case is similar to that of galactonic acid and its lactone (p. 774).
Double Lactones.
With the
dibasic acids derived from d-
and
1-mannose, the peculiarity of double lactone f formation is observed. These very characteristic compounds crystallize out with 2 molecules of
water, which can be eliminated by drying over concentrated sulphuric acid. Aqueous solutions of the double lactones are at first neutral,
but become acid upon standing; the aqueous solutions have also. the peculiarity of strongly reducing Fehling's solution, this being probably due to an aldehydic rearrangement of the dilactone molecule in presence of free alkalies.
and double lactones agree perHudson's hypothesis (p. 774) according to which the character of rotation depends upon the position of the lactone ring. The structure of the double lactone of d-mannosaccharic acid is
rotations of the lactone acids
fectly with
The
shown
as follows:
Tollens and Sohst,
t Kiliani, Ber., 20,
Chem. Ztg., 11, 99, 1178j Ann., 246, 341; Fischer, Ber., 24, 539.
1.

The property
781
of undergoing transformation to other isomers upon with pyridine at 140, noted for the monobasic acids (p. 775), heating also exists with the dibasic acids. Mucic acid has been converted in this * way by Fischer into the isomeric compound allomucic acid.
COOH
COOH
HCOH HOCH HOCH HCOH COOH

Mucic acid
HOCH HOCH HOCH HOCH COOH

Allomucic acid
As with the monobasic
acids the groups adjoining are the parts of the molecule affected in this reaction.
HCOH
COOH radicals
Dehydration of Dibasic Acids of Hexoses.
acteristic of the dibasic acids of the hexoses is the ease
noteworthy charwith which they
undergo dehydration, upon heating to 150 C. with concentrated hydrochloric acid, hydrobromic acid, sulphuric acid or other dehydrating The agent, with formation of the unsaturated dehydromucic acid.
reaction
is
illustrated graphically as follows:
H
:
H
c ion";
/ \ COOH O fcT;
H
_
c
/ \
HO
ic
/ \
.
/\
O
HOOC
HOOC
COOH
Water.
|"H"
Hexose dibasic acid
Dehydromucic acid
Dehydromucic Acid.
The best dehydrating agent to use for the above
and hydroreaction, according to Fischer,* is a mixture of hydrochloric Fischer considers the dehydromucic acid reaction the bromic acids.
best of
all
methods
Dehydromucic and Yoderf: 2 to 5 mgs.
acid
for detecting a dibasic acid of the hexose type. is best recognized by the reaction of Tollens
of substance are carefully heated with 2 c.c. concentrated sulphuric acid and 1 to 4 mgs. of isatin at 145 to 155 C. When the test is made with pure dehydromucic acid the solution will be colored a strong violet blue; with the dibasic hexose acids (mucic, mannosaccharic, etc.), the solution takes on more of a green
saccharic,
color and shows before the spectroscope two characteristic absorption bands near the a and (3 lines of strontium.
*
Her., 24, 2136.
t Ber., 34, 3448.
782
SUGAR ANALYSIS
splits off
Dehydromucic acid upon heating

mucic acid which
is
C02
(p.
and
yields pyro-
the acid derivative of furfural
374).
H
C
H -C
H
C
-H C
O
\ / \ O COOH
Pyromucic acid
\ / \
CHO
Furfural
Chitonic and Isosaccharic Acids.

in structure are the saturated
Resembling dehydromucic acid monobasic and dibasic acids derived

itself
from
chitose,
which
is
probably also
a saturated furfuran de-
rivative.
HOC
-H COH
CH
CHO
O
HC
HOH C
2
/\/\
Chitose
H HOG HC
HOH C
2
H -COH
CH
O
HOC
HOOC
-H COH
CH
O
HC
/ \ / \
/ \ / \
COOH
COOH
Chitonic acid
Isosaccharic acid
The dibasic acids of the sugars Hydrazides of Dibasic Acids. the same the monobasic as derivatives; the second hydrazides yield carboxyl group enables them however to fix an additional molecule of phenylhydrazine. Many of the dibasic acids give, in fact, two classes The acid hydrazides of compounds, the acid and double hydrazides. are precipitated usually with phenylhydrazine in the cold and the double
hydrazides by heating. The following formulae illustrate the configuration of the acid and double hydrazides:
COOH
(CHOH) 4
I
OC-N-N-C H
6
H H
OC-N-N-C H5
6
H H
(CHOH) 4
|
OC-N-N-C H
6
H H
Acid phenylhydrazide
Double phenylhydrazide.
The
acid hydrazides are colorless
compounds
easily soluble in hot
water, while the double hydrazides are usually of a pale yellow color and only slightly soluble in hot water.
are reduced
Reduction of Dibasic Acids. The lactones of the dibasic acids by sodium amalgam, following the same method described on p. 776, and yield in succession the lactones of the monobasic acid, the sugars and the corresponding alcohols.
dibasic
Add. An interesting intermediary step between the and monobasic acids, noted in the reduction of the lactones of saccharic and mucic acids, is the production of an aldehyde acid. In
d-Glucuronic
783
the case of saccharic acid monolactone, for example, Fischer and Piloty obtained as an intermediary reduction product d-glucuronic acid.
COOH
COOH
CHOH
)H
O
I
Saccharic acid monolactone
Uo
CHOH CHOH
I
CHOH CHOH H = CHOH CHOH

2
mo
d-Glucuronic acid
upon
d-Glucuronic acid occurs naturally in the urine and yields furfural distillation with hydrochloric acid; its properties, reactions, and close relationship to the pentoses are referred to elsewhere (p. 375). The successive steps in the reduction of different lactones of the
dibasic acids are given as follows:

Dibasic acid lactone.
784
SUGAR ANALYSIS
is
low solubility in cold water and this property

identification of these acids.
made
use of in the
There are a large number of interesting double salts of the dibasic Several dibasic acids, acids but only a few of these can be mentioned. as tartaric, saccharic and mucic, give double compounds with potassium
and antimony
emetic,
is
oxide.
Of these potassium antimonyl
tartrate, or tartar
given as illustration:
COOK
J,
HOH
COO-Sb=O.
ammonium tartrates have a was owing to the work of Pasteur upon these salts that the science of molecular asymmetry and the methods for analyzing racemic mixtures had their first beginning. The problem of separating the dextro- and levo-rotatory components of an optically inactive racemic mixture was in fact first solved by Pasteur; as the methods established by him are still the ones most generally
Of other double
salts
the sodium
special historical interest, since it
employed, this particular branch of sugar analysis may be treated best in connection with a review of Pasteur's work upon tartaric acid.
THE ANALYSIS OF RACEMIC MIXTURES

Tartaric acid
may
be said to exist under four different forms; the
structural formulae of these are represented as follows:
COOH
COOH
COOH
COOH
COOH
HCOH HOCH COOH

Dextro- or
d-tartaric acid
I
HOCH HCOH COOH

Levo- or
1-tartaric acid
HCOH HCOH COOH

Meso- or
i-tartaric acid
HCOH HOCH COOH
HOCH HCOH COOH

.
Racemic or
d, 1-tartaric acid
II
(inactive) III
(inactive)
IV
The d- and 1-components of a racemic * mixture usually resemble one another in melting point, solubility, specific gravity, chemical affinity, and all other properties except specific rotation; the racemic substance
may differ, however, from its components in crystalline form, melting point, solubility, and other characteristics. In other words a racemic compound may behave not as a mixture, but as a simple subitself
The word racemic is derived from the Latin for tartaric acid, acidum racemicum, where the phenomenon was first noted.

stance;
785
it is this peculiarity which renders the separation of the two optically active antipodes in a racemic mixture a matter of such diffi-
culty.
laboratory operations where optically active substances d- and 1-isomers are produced in equal amounts; many the are formed, of instances The optical activity escape notice for this very reason.
In
many
possibility of separating
cally active
an optically inactive compound into two opti-
components should therefore always be considered. Separation of Racemic Mixtures by Differences in Crystalline Form. It was observed by Pasteur* in 1848 that when a solution of racemic
Fig. 200.
Showing opposite hemihedrism of d-tartaric and
of crystals of the
1-tartaric acids.
sodium-ammonium salts
acid which
one-half with
tions,
neutralized, one-half with sodium hydroxide and ammonia, was allowed to evaporate under certain condiseparate crystals were obtained of the d- and 1- double salts. The
had been
two
classes of salts
were similar in
all
respects except in the position of
(shown in black, Fig. 200). In one set of crystals, for example, the hemihedral faces were always at the right of the surfaces d and e, when the latter were uppermost, and in the other set of The relationship between the two crystalcrystals always at the left. line forms was exactly like that between one crystal and its mirror image, where one form cannot be brought into coincidence with the
their hemihedral faces
other
by any method
of turning the crystal.
separately the two sets of hemihedral crystals, Pasteur obtained in one case a solution which rotated the plane of polarized
By dissolving
light to the right,
and
in the other case a solution

left.
which rotated the

effected of
plane of polarized light to the
Separation was thus
* For a full account of Pasteur's researches upon the tartaric acids see his Re"cherches sur la dissymmetric moleculaire des produits organiques naturels," Paris; also his original papers, Compt. rend., 26, 535; 27, 401; 32, 110; 36, 180; 36,
"
26; 37, 110, 162; etc.
786
SUGAR ANALYSIS
of
The its two optically active components. hemihedrism was explained by Pasteur as due to an asymmetric arrangement of the atoms within the molecule, the grouping in one compound being exactly the reverse of that in the other. If the d, 1-sodium ammonium tartrate crystallizes out at a high temperature only the non-hemihedral crystals of the racemate are obtained. The transition point between separation of racemate and that of the hemihedral crystals of d- and 1-tartrate is 28 C. and it is only under this temperature that separation of the two salts can be effected by the
the inactive racemic salt into
phenomenon
difference in crystalline form. Separations of racemic mixtures into their optically active components by differences in crystalline form have been made upon other
optically inactive lactone of d, 1-gulonic acid, for example, crystallizes, according to Haushofer,* in rhombic crystals with hemihedral faces; by selecting the forms of opposite hemihedry
sugar derivatives.
The
the d- and 1-lactones are obtained of opposite specific rotation. This means of separation is not, however, generally applicable, and recourse
is
usually
made
to other methods.
Racemic Mixtures by Combination with Other OptiActive This second method of separating racemic cally Compounds. mixtures is also due to Pasteur, who discovered that when a hot aqueous
Separation of
solution of d, 1-tartaric acid
different cinchona bases the quinine
was saturated with equivalent amounts of and quinicine salts of d-tartaric
acid crystallized out before the corresponding compounds of 1-tartaric acid, while the cinchonine and cinchonicine salts of 1-tartaric acid
separated before the corresponding compounds of d-tartaric acid. This method of separating racemic mixtures has been greatly extended since the time of Pasteur and has been applied to many different classes
of
compounds. In many operations where sugar acids are formed, both optical antipodes are produced, the inactive racemic mixture of the d- and 1-acids behaving very much as a simple acid and yielding upon evaporation an optically inactive lactone. The salts of the alkaloids have been of great service in separating the
dsalt of
and 1-components of different inactive sugar d-mannonic acid,f for example, is soluble
acids.
The
strychnine
in hot absolute alcohol,
while the strychnine salt of 1-mannonic acid is insoluble. If the latter is filtered off, dissolved in water and treated with barium hydroxide solution, the strychnine is precipitated and a soluble barium salt of 1-mannonic acid formed. The solution is filtered, shaken out with ether to
*
Ber., 24, 530; 26, 1027.

in exact
787
remove any remaining strychnine and then treated with sulphuric acid amount to precipitate all barium sulphate. The latter is filtered off and the filtrate evaporated when the 1-mannonic acid will In the same manner d-galactonic acid crystallize out as a lactone. salt of low solubility) has been separated from 1-galactonic (strychnine
acid.
principal alkaloids used for separating racemic mixtures of acids the are cinchona bases, quinine, quinidine, cinchonine and cinchonidine; the strychnos bases, strychnine and brucine; and the opium base,
The
morphine.
principle of this method has also been employed in separating racemic mixtures of sugars by means of optically active hydrazines
(p. 361).
The
Separation of Racemic Mixtures by Selective Fermentation. This third method of separating racemic mixtures is also due to Pasteur and is based upon the difference in susceptibility of the d- and
1-components to attack by different ferments and moulds. Pasteur noted that inactive solutions of ammonia d, 1-tartrate after inoculation
with spores of Penicillium glaucum (in presence of slight amounts of mineral salts to act as nutrients) became strongly levorotatory. This
was explained by the fact that the d-tartaric acid was fermented by the mould, the 1-tartaric acid remaining unaffected. Pasteur's third method of resolving racemic compounds has also been greatly extended and has been employed with success in sepa-
Thus by means of yeast rating mixtures of d, 1-sugars and acids. Fischer was able to ferment the d-sugar in d, 1-glucose, d, 1-mannose,
d, 1-galactose
and
d, 1-fructose,
and obtain the 1-sugar
in
a pure con-
dition.
For separating the sugar acids, Penicillium glaucum, first used by PasOf the acids fermented by this mould, teur, is still largely employed. be mentioned may d-tartaric, d-glyceric, d-mannonic, and d-glutaminic The of these compounds not being attacked. the 1-isomers acids, selective influence of a mould, yeast, or other organism is not confined, however, to the members of a single d- or 1-series as might be inferred from the examples mentioned. Thus with the ammonium salt of d, 1lactic acid (fermentation lactic acid), the 1-lactic acid is fermented by Penicillium glaucum and the d-compound left behind in solution.
APPENDIX OF SUGAR TABLES
INTRODUCTION
THE
following tables, which have been selected to
accompany various
methods described
in the author's Handbook of Sugar Analysis," have been grouped together for convenience as a separate Appendix.
"
This arrangement was

of the text
made
partly to prevent breaking the continuity
by
the introduction of lengthy tables and partly to permit
the publication of the Appendix as a separate book for the convenience

of those
who have occasion to make use of special tables in the laboratory.
the author has
Knowing the very diverse preferences of individual sugar chemists, made a rather wide selection from the more commonly
Limitations of space have obliged him,
used copper reduction tables.

however, to leave out
his excuse for
many
tables of recognized merit
and
this
must be
any
errors of omission.
LIST OF TABLES
TABLE
1.
PAGE
Specific
Gravity of Sucrose Solutions at ^5- C.
20
(Kaiserliche
Normal1
Eichungs-Kommission)
2.
Temperature Corrections for Changing Percentages of Sugar by Specific

Gravity to True Values at 20
C
1
5
7
1
'
'i
3.
Specific
Gravity of Sucrose Solutions at
.o
C. with Corresponding
Degrees Brix and Baume",

4.
Table for Correcting Readings of Brix Hydrometers at Different Temperatures to 17.5
16
5.
Main's Table for Determining Water in Sugar Solutions by Means of the Abbe Refractometer at 20 C
Stanek's Correction Table for Determining Water in Sugar Solutions by Means of the Abbe Refractometer when Readings are Made at Other
17
6.
Temperatures than 20
7.
21
Geerligs's Table for
by the Abbe Refractometer

8.
Determining Dry Substance in Sugar House Products at 28 C
22
Hiibener's Table for Determining Percentages by Weight of Sucrose in Sugar Solutions from Readings of the Zeiss Immersion Refractometer
Kruis's Table for Determining Glucose
Allihn's Table for
Pfliiger's
24
9.
by Reischauer's Method
27 30
33
35
10. 11. 12. 13. 14. 15. 16.
Determining Glucose
Table for Determining Glucose

for
Koch and Ruhsam's Table
Determining Glucose in Tanning Materials
Meissl's Table for Determining Invert Sugar
38
40 42
Wein's Table for Determining Maltose

Soxhlet and Wein's Table for Determining Lactose
Woy's Table for Determining Glucose, Fructose, Invert Sugar, Lactose and Maltose by Kjeldahl's Method
Brown, Morris and Millar's Table Invert Sugar
for
44
17.
Determining Glucose, Fructose and

62
63
18.
Defren's Table for Determining Glucose, Maltose and Lactose
19. Munson and Walker's Table for Determining Glucose, Invert Sugar Alone, Invert Sugar in the Presence of Sucrose (0.4 gram and 2 grams Total
Sugar), Lactose
and Maltose
vii
66
viii
SUGAR TABLES
PAGE
Glucose,
Galactose,
TABLE 20. Bertrand's Table for Determining Invert Sugar, Maltose and Lactose
21.
79
Herzfeld's Table for Determining Invert Sugar in Sugar not to Exceed 1.5%)
Raw
Sugars (Invert
81
22.
23.
Krober's Table for Determining Pentoses and Pentosans

Tollens, Ellet
83
and Mayer's Table
for
Determining Methylpentoses and

89
Methylpentosans
24.
Formulae, Descriptions, Melting Points and Solubilities of the Principal Hydrazones and Osazones of the Sugars
Reciprocals of
90
101
25.
Numbers from
to 100
SUGAR TABLES
C^ O SO oooo o
T
I
to OS CO
i
ISI-H
C^l
1
^* OO
"ft
Tf CO
tO
i
i
l>-
d O ii
tO C^l f^ CO to C3 <N to
i
i
OO ^H CO t^* OS C^l t~- to b!>
OQO
(N "^
>
C^ <N C^ CO CO
-rfi
"*
T)-I
to
CO
'
'
rt< OS IO iO OS iO OS GO t^ CO CO
Ô
i-H
CO C^ tO t^ (N CO CO t^
'
-H
kO OS CO t^
,00
OS CO
i
i
8!
to
t^- r
OS OS
dOOOOO'
Tt<
8
COCOb
!> t^
03 OQ (N CO CO
-^ Tf to tO
C^l
CO OS OO
T-H
;SS
SS
T-H
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SUGAR TABLES
C^C^COÔ
t^.C^brHCO
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SUGAR TABLES
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C^
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t>T-H
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Tf
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1
T-H CO T-H T-H
00 lO
Si
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CO OS CD CO
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to to to
l> CO <N (N T-H OS T-H CO tO CO CO T-H t^
r^t-T-tQ'*
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Ttl
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to
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l>
SUGAR TABLES
TABLE*
2.
TEMPERATURE CORRECTIONS FOR CHANGING PERCENTAGES OF SUGAR BY SPECIFIC GRAVITY TO TRUE VALUES AT 20 C.
SUGAR TABLES
TABLE*
3.
SPECIFIC
GRAVITY OF SUCROSE SOLUTIONS AT
17.5
C.
17.5
WITH CORRESPONDING
DEGREES BRIX AND BAUME

Per cent sucrose by weight or degrees
Brix.
SUGAR TABLES
TABLE
Per cent sucrose by weight or degrees Erix.
3.
(Continued.)
SUGAR TABLES
TABLE
3.
(Continued.)
Per cent sucrose by weight or degrees Brix.
SUGAR TABLES
TABLE
3.
(Continued.)
10
SUGAR TABLES
TABLE
3.
(Continued.)
SUGAR TABLES
TABLE
3.
11
(Continued.)
12
SUGAR TABLES
TABLE
3.
(Continued.)
SUGAR TABLES
TABLE
Per cent sucrose by weight or degrees
Brix.
13
3.
(Continued.)
14
SUGAR TABLES
TABLE
3.
(Continued.)
SUGAR TABLES
TABLE
Per cent sucrose by weight or
3.
15
(Concluded.)
16
SUGAR TABLES
TABLE*
4.
TABLE FOR CORRECTING READINGS OF BRIX HYDROMETERS AT DIFFERENT TEMPERATURES TO 17.5C.
SUGAR TABLES
TABLE*
MAIN'S TABLE FOR DETERMINING
5.
17
SUGAR SOLUTIONS BY MEANS OF THE ABBE REFRACTOMETER.

IN
WATER
Refractive index at 20 C.
18
SUGAR TABLES
TABLE
5.
(Continued.)
SUGAR TABLES
TABLE
5.
19
(Continued.)
20
SUGAR TABLES
TABLE
5.
(Continued.}
SUGAR TABLES
TABLE
5.
21
(Concluded.")
22
SUGAR TABLES
TABLE*
7.
GEERLIGS'S TABLE FOR DETERMINING DRY SUBSTANCE IN SUGAR-HOUSE PRODUCTS. By the Abbe Refractometer, at 28 C.
R6fric~
SUGAR TABLES
TABLE
7.
23
(Concluded.)
CORRECTIONS FOR TEMPERATURE.
24
SUGAR TABLES
TABLE*
8.
HUBENER'S TABLE FOR DETERMINING PERCENTAGES BY WEIGHT OP SUCROSE IN SUGAR SOLUTIONS FROM READINGS OF THE ZEISS IMMERSION REFRACTOMETER.
ijj
SUGAR TABLES
TABLE
sti
25
8.
(Continued.)
26
SUGAR TABLES
TABLE
8.
(Concluded).
99
20.23
.25 .27 .29
.31
100.0 20 44
.1
101.0 20 66
.1
102.0 20.87
.1
103.0 21.08
.1 .2 .3 .4 .5 .6 .7
104.0 21.29
.1
105
1
21 51
2
.34 .36 .38 .40 .42
.2 .3 .4 .5 .6 .7 .8 .9
.47 .49 .51 .53 .55 .57 .59 .61
.2
.3 .4 .5 .6 .7
.68 .70 .72 .74 .76 .78 .80 .82 .85
.89
.91
.2
.93 .95 .97
21 00
.02 .04
.10 .13 .15 .17 .19 .21 .23 .25 .27
.2
.31 .34 .36 .38 .40 .42 .44 .47 .49
4 5 6
7
.9
.53 .55 .57 .59 .61 .63 .66 .68 .70
106.0
21.71
SUGAR TABLES
TABLE*
Fehling's
solution.
27
9.
KRUIS'S TABLE FOR DETERMINING GLUCOSE BY REISCHAUER'S METHOD.
28
SUGAR TABLES
TABLE
9.
(Continued.)
Fe'iling's solution.
SUGAR TABLES
TABLE
Fehling's
solution.
29
9.
(Concluded.)
30
SUGAR TABLES
TABLE*
10.
ALLIHN'S TABLE FOR DETERMINING GLUCOSE.

Cop-
t&i
SUGAR TABLES
TABLE
Copper
(Cu).
31
10.
(Continued.)
32
SUGAR TABLES
TABLE
10.
(Concluded.)
Ogpe,
SUGAR TABLES
TABLE*
11.
33
PFLUGERS TABLE FOR DETERMINING GLUCOSE.

Glucose.
34
SUGAR TABLES
TABLE
11.
(Concluded.)
Glucose.
SUGAR TABLES
TABLE*
Copper.
(Cu).
35
12.
KOCH AND RUHSAM'S TABLE FOR DETERMINING GLUCOSE IN TANNING MATERIALS.
36
SUGAR TABLES
TABLE
12.
(Continued.)
Copper.
(Cu).
SUGAR TABLES
TABLE
Copper.
(Cu).
37
12.
(Concluded.)
38
SUGAR TABLES
TABLE*
13.
MEISSL'S TABLE FOB DETERMINING INVERT SUGAR.

Copper.
(Cu).
SUGAR TABLES
TABLE
Copper.
(Cu).
39
13.
(Concluded.)
40
SUGAR TABLES
TABLE*
14.
WEIN'S TABLE FOR DETERMINING MALTOSE.

Cop(Cu).
SUGAR TABLES
TABLE
Cop(Qu).
41
14.
(Concluded.)
42
SUGAR TABLES
TABLE*
15.
SOXHLET AND WEIN'S TABLE FOR DETERMINING LACTOSE.

<
&T
SUGAR TABLES
TABLE
c
(
43
15.
(Concluded.)
44
SUGAR TABLES
TABLE*
16.
WOY'S TABLE FOR DETERMINING GLUCOSE, FRUCTOSE, INVERT SUGAR, LACTOSE AND MALTOSE BY KJELDAHL'S METHOD.
15 c.c. Fehling's Solution.
Cupric
oxide (CuO).
SUGAR TABLES
TABLE
Cupric
oxide (CuO).
16. (Continued.) 15 c.c. Fehling's Solution.
45
46
SUGAR TABLES
TABLE
16. (Continued.) 15 c.c. Fehling's Solution.
Cupric
oxide (CuO).
SUGAR TABLES
TABLE
16.
47
(Continued.}
15 c.c. Fehling's Solution.

Cupric
oxide (CuO).
48
30
Cupric
oxide
SUGAR TABLES
TABLE
c.c.
16. (Continued.) Fehling's Solution.
(CuO).
SUGAR TABLES
TABLE
30
Cupric
oxide (CuO).
c.c.
49
50
SUGAR TABLES
TABLE
30
c.c.
Cupric
oxide
(CuO).
SUGAR TABLES
TABLE
30
Cupric
oxide (CuO).
c.c.
51
16.
(Continued.)
Fehling's Solution.
52
SUGAR TABLES
TABLE
30
c.c.
16.
(Continued.)
Fehling's Solution.
Cupric
oxide (CuO).
SUGAR TABLES
TABLE
50
Cupric
oxide (CuO).
c.c.
53
16.
(Continued.)
Fehl ing's Solution.
54
SUGAR TABLES
TABLE
50
c.c.
Cupric
oxide
(CuO).
SUGAR TABLES
TABLE
50
Cnpric oxide (CuO).
c.c.
55
56
SUGAR TABLES
TABLE
50
c.c.
Cupric
oxide (CuO).
SUGAR TABLES
TABLE
50
Cupric
oxide (CuO).
c.c.
57
16.
(Continued.)
Fehling's Solution.
58
SUGAR TABLES
TABLE
16. (Continued.} 50 c.c. Fehling's Solution.
Cupric
oxide (CuO).
SUGAR TABLES
TABLE
50
Cupric
oxide (CuO).
c.c.
59
16.
(Continued.)
Fehling's Solution.
60
SUGAR TABLES
TABLE
50
c.c.
16.
(Continued.)
Fehling's Solution.
Cupric
oxide (CuO).
SUGAR TABLES
TABLE
50
Copper
oxide
16.
61
(Concluded.)
c.c.
Fehling's Solution.
(CuO).
62
SUGAR TABLES
TABLE*
17.
BROWN, MORRIS AND MILLAR'S TABLE FOR DETERMINING GLUCOSE, FRUCTOSE AND INVERT SUGAR.
Milligrams
of sugar.
SUGAR TABLES
TABLE*
Cupric
oxide.
63
18.
DEFREN'S TABLE FOR DETERMINING GLUCOSE, MALTOSE AND LACTOSE.
(CuO).
64
SUGAR TABLES
TABLE
18.
(Continued.)
Cupric oxide
(CuO)~
SUGAR TABLES
TABLE
Cupric oxide
18.
65
(Concluded.')
66
SUGAR TABLES
TABLE*
19.
MUNSON AND WALKER'S TABLE FOR DETERMINING GLUCOSE, INVERT SUGAR ALONE, INVERT SUGAR IN THE PRESENCE OF SUCROSE (0.4 GRAM AND 2 GRAMS TOTAL SUGAR), LACTOSE AND MALTOSE.
SUGAR TABLES
TABLE
19.
67
(Continued.')
68
SUGAR TABLES
TABLE
19.
(Continued.}
SUGAR TABLES
TABLE
5
19.
69
(Continued.)
70
SUGAR TABLES
TABLE
19.
(Continued.)
SUGAR TABLES
TABLE
I
19.
71
(Continued.)
72
SUGAR TABLES
TABLE
19.
(Continued.)
SUGAR TABLES
TABLE
19.
73
(Continued.)
74
SUGAR TABLES
TABLE
19.
(Continued.)
SUGAR TABLES
TABLE
i
19.
75
(Continued.)
76
SUGAR TABLES
TABLE
19.
(Continued.}
SUGAR TABLES
TABLE
i
19.
77
(Continued.}
78
SUGAR TABLES
TABLE
19.
(Concluded.}
SUGAR TABLES
TABLE*
20.
79
BERTRAND'S TABLE FOR DETERMINING INVERT SUGAR, GLUCOSE, GALACTOSE, MALTOSE, AND LACTOSE.
Milligrams of
sugar.
80
SUGAR TABLES
TABLE
20.
(Concluded.)
Milligrams of
sugar.
SUGAR TABLES
TABLE*
21.
81
HERZFELD'S TABLE FOR DETERMINING INVERT SUGAR IN SUGAR NOT TO EXCEED 1.5%.)
Oogjr.
RAW
SUGARS (INVERT
82
SUGAR TABLES
TABLE
21.
(Concluded.)
Copper.
(Cu).
SUGAR TABLES
TABLE *
Furfural phloroglucide.
83
22.
KROBER'S TABLE FOR DETERMINING PENTOSES AND PENTOSANS.
84
SUGAR TABLES
TABLE
22.
(Continued.)
SUGAR TABLES
TABLE
85
22.
(Continued.)
86
SUGAR TABLES
TABLE
22.
(Continued.)
SUGAR TABLES
TABLE
87
22.
(Continued.)
88
SUGAR TABLES
TABLE
22.
(Concluded.)
SUGAR TABLES
TABLE*
23.
89
TOLLENS, ELLET, AND MAYER'S TABLE FOR DETERMINING METHYLPENTOSES
AND METHYLPENTOSANS.
Methylfurfural phloroglucide.
90
SUGAR TABLES
"-< -
--<
-^
03
03
~
03
03
0)
03
03
"
03
CO
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t
I
O
I
t>-
OS
O O O5
SUGAR TABLES
91
o o
-a
00 ~~
,
o
-*
<M <N <N CO OO T-H <M t iO C^ <M C^
O
1
-CO
^ t>
T
^00
icTcOCD
l-H
Tt< 1-1 I-H
Q CO CO coîScoii t^ CO CD CD
CO
l-H T-H i-H T-l C<l
>-H
CO CO
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J-H
0)
3
^
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%
053
bC
t,' bfi
02
CO
02
00
02
02
<u
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oo-dô^Sooo 73
K G S c S S*G O
occoobobbb
o o
4
S S 3
S^
oooodooooo
2 c5d 2 c5du o 2 2 dodo o c5d

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92
SUGAR TABLES
WWW
o o XXX O O
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05
SUGAR TABLES
3
a
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03 oi
93
a
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a
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C<l C^l C<I
C<l i-H
CO
s
o> co
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S3
O
02
fa
J2
tj_ t_i
(
RM X ^^ ^_2^^ ^^ ^^^
02
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03
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CO
rt
z; g G fl e ^ ^ 15 ^ G fl O O g O o Or^r: z3:
:!;
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C2
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fa
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e e ^ ^ o o
c a e ^ o o o
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94
SUGAK TABLES
J=J^.S ^ ^
^-^^.S^
aw
00
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^000
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i^^^^^ô^?' o o >,o o
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'
s s s
ill
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'03'03'03
G G
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ooo
^
O-fôjQj' 'i
Isolactose
Turanose
Melibiose
Gentiobiose
Cellose
Glucosidogalactose Galactosidogalactos Mannatrisaccharide
rf,Z-Glycer d-Erythro
Apiose
03
cl
=1
ol
03
:'::'
OJ OJ
03
^3 ^5 J3 ^3 45
0!J
02
03
02
02
02-
QQQQQQQH
SUGAR TABLES
95
alcoho alcoho
Jg 11
Sligh Alco
Pyri
Pyridine+methylPyridine+methyl
<N
1
OO
r-ICOOi-HlOO<N<0<N
LOGO
1
00 OO
rHi-^t-HlOr 00 00
OSOiOtÔS Ir
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1
03
1
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02
03
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02
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03
a a a a a a
,2,2
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03
03
03
03
03
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03
a a a
03
03
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02
as
03
03
03
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0,2,2
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p5
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25.
1
RECIPROCALS OF NUMBERS FROM

Number.
TO 100.
INDEX
Abbe
reflactometer, 53-61.
adjustment, 59-61.
compensator, 57, 58.

Geerligs's table for, 65; Appendix, 22. illumination, 58.
Main's table
for, 64; Appendix, 17. temperature regulation, 58, 59. theory of, 53-57.
"Absatz" method
Absorption error of
of
of Tollens, 344, 345.
bone black, 220, 221, 284, 285

7, 8.
moisture by raw sugars,
spectra (see Spectra). Accessories of polariscopes, 146-171.
Acetaldehyde, reaction with sugars, 368. sugar alcohols, 766 Acetals of sugar alcohols, 766. Acetates of lead, 207, 208 (see Lead).
Acetic acid, inverting power, 273, 663. method for decomposing saccharates, 250. anhydride, reaction with sugars, 369.
Acetol, 536.
Acetylcarbinol, 536.
Acetylene lamps, 152. Acetylmethylcarbinol, 537. Achroodextrin, 577, 686. Acidity of sugar products, determination, 496, 497. Acids, color reactions with sugars, 340, 341.
influence
on activity of
diastase, 691.
invertase, 671.
pancreatin, 694. Clerget factor, 269. rotation of sugars, 185, 186.

inverting power, 662-666. organic, for invert polarization, 273.
products from heating sugars with, 340, 341.

relation of affinity, inverting
power and conductivity,
Acids of the sugars, 529, 772-787. dibasic, 778-787.

dehydration, 781.
double lactones, 780.

formation, 778.
xiii
xiv
Acids of the sugars:
dibasic,
INDEX
hydrazides, 782. lactone acids, 779.
nomenclature, 778-779.
properties, 779.
reduction, 782, 783.

salts, 783, 784.
monobasic, 772-778.
hydrazides, 777.
lactones, 773, 774 (see Lactones). molecular rearrangement, 775. nomenclature, 773.
oxidation, 778. salts, 777, 778.

synthesis, 772, 773.
Acorn sugar (see Quercite). a- and -Acrose, 623.

Adonite, dibenzal, 770.
occurrence, 559. oxidation to d, 1-ribose, 559.
ketopentose, 562.
properties, 767. Affining, 646.
Affinity and inverting power of acids, 663. Agar-agar, preparation of d-galactose from, 603. Alcohol (ethyl), digestion methods
(see
Sugar beets),
extraction methods (see Sugar beets), influence on rotation of sugars, 181, 182.
activity of invertase, 674, 675.
lamps, 152.
precipitate, determination in fruit products, 520. honey, 521. use of in purification of sirups, 550. Alcoholic fermentation, 581, 582, 604, 619, 651, 701, 702, 714 738. Alcohols, reaction with sugars, 367. Alcohols of the sugars, 529, 530, 764-772.
compounds with metals, 765. formation during fermentation, 764, 765. nomenclature, 766. oxidation by bacteria, 771, 772.
chemical means, 770, 771.
properties, 765-770. reactions with
acetaldehyde, 766.
benzaldehyde, 766, 769, 770. borax and boric acid, 765.
formaldehyde, 766.
rotation, 765-768.
influence of boric acid on, 765, 766. molybdic acid on, 766.
INDEX
Alcohols of the sugars.
rotation, influence of tungstic acid on, 766. synthesis, 764.
XV
table of classification, etc., 767, 768.
Aldehyde reactions
of sugars, 333-387, 527.
Aldehydes, reaction with sugars, 368. Aldoses, conversion into ketoses, 355,
distinguishing from ketoses, 340, 354, 363, 380. group, 527.
oxidation with bromine, 363.
Aldoheptoses, 633-637. Aldohexoses, 570-612.
Aldopentoses, 545-560. Aldotetroses, 540-542.

Aldotrioses, 538. Alkalies, action upon d-galactose, 603, 604. d-glucose, 586, 587.
lactose, 712, 713.
maltose, 701.
reducing sugars, 303, 339, 340. color reactions with sugars, 339. influence on activity of invertase, 671.
diastase, 691.
pancreatin, 694
mutarotation, 190.
rotation of sucrose, 183.
products from heating sugars with, 339, 340. saccharates of, 676, 677.
Alkaline earths, influence on rotation of sucrose, 183. saccharates of, 677.
Alkalinity of sugar products, determination, 496, 497. Alkaloids, use in resolving d, 1-acids, 786, 787.
Allen's
Allihn's
method for determining glucose, maltose and dextrin, 486-488 method for determining glucose, 403; Appendix, 30.
application to other sugars, 420, 421. modification by Koch and Ruhsam, 420; Appendix, 35.
Pfliiger,
419; Appendix, 33.
Allylphenylhydrazine, 346.
Allomucic acid, 781.
D and [a]j, meaning of symbols, Alum as a clarifying agent, 223.

[a]
172.
Alumina cream, preparation,
Aluminum hydroxide
Amines, Amino compounds, influence on Clerget factor, 270.
222, 223. for clarifying, 222, 223. reaction with sugars, 367.
Amino
Ammonium
sugars, 751-754. nitrate method for decomposing saccharates, 251.
Amygdalin, 572. Amygdalinbiose, 730. Amylases (see under Conversion of starch).
xvi
Amylocellulose, 688.
INDEX
Amylodextrin, 577, 686, 706. Amylodextrinase, 686. Amyloglucase, 691. Amylomaltase, 686. Amylopectin, 688. Amylose, 688. Amylphenylhydrazine, 346. d-Amylphenylhydrazine, use in resolving racemic sugars, 362.
Analytical balance, 39, 162. Analyzer, 82-84.
Angular rotation, calculation to saccharimeter degrees, 145.

determination of sugars from, 194, 195. Aniline acetate test for artificial invert sugar, 620.
furfural, 374-375.
methylfurfural, 377.
Animal
test-paper, 375. cellulose, 579.
gum, 579.
Antiarin, 569. Antiarose, 569.
Apiin, 544.
hydrolysis to glucoapiose, 643.

Apiose, 544, 644. Apparatus, care of polariscopic, 169-171.
Araban, determination, 450-452; Appendix, 83

hydrolysis to 1-arabinose, 548.
occurrence, 546-548.
properties, 546, 547. Arabic, gum, 547. Arabinic acid, 547, 601.
(see also
under Pentosans).
d-Arabinose, 545. formation from galactoarabinose, 644.
1-menthylhydrazone, 362, 545, 551. synthesis of glucosamine from, 754.
from d-glucose, 365.

1-Arabinose, 546-551.
absorption spectra with a-naphthol, 379.

phloroglucin, 384.
resorcin, 381.
calorific value, 319.
conversion to 1-glucose, 592.
1-mannose, 597.
1-ribose, 777.
determination, 450-452; Appendix, 83 (see also under Pentoses). as diphenylhydrazone, 469.

in presence of fructose, 482.
xylose, 482.
fermentation, 550, 551. action of different yeasts, 714.
INDEX
1-Arabinose, formation
xvii
548.
by hydrolysis from arabans,
diarabinose, 643.
mutarotation, 187.
occurrence, 546. preparation from cherry gum, 548-550.
properties, 550, 551. reducing ratio to glucose, 421. specific rotation, 174-192, 550.
tests, 551.
value of Ventzke degree, 200, 201. yield of furfural from, 449. d, 1-Arabinose, 551. resolution of, 362.
d-Arabite, 545, 557, 767. 1-Arabite, 551, 767.
monobenzal, 770Arabogalactans, 599.
d-Araboketose, 561.
1-Araboketose, 561. d-Arabonic acid, 545.
oxidation to d-erythrose, 540.
1-Arabonic acid, 551. conversion to 1-ribonic acid, 559, 775. rotation of lactone, 551, 774. oxidation to 1-erythrose, 541
.
Arbutin, 571.
Armstrong on enzymic synthesis
of sugars, 704, 705.
Arrhenius's hypothesis of inversion, 664. viscosity equation, 310.

406. Asbestos, preparation for filter-tubes and Gooch crucibles, Ash, analysis of for determining origin of sugars, 519. determination by direct incineration, 495. ignition with sulphuric acid, 495.
in
commercial dextrins, 509.

sugar products, 495.
of
maple sugar, composition, 519. muscovado sugar, composition, 519.
496. preparation for quantitative analysis, 495,

soluble
and
insoluble, 495.
246. Asparagine error in sugar beet analysis, 245, Aspergillus niger, action
upon gentianose,
743.
lactose, 715.
melezitose, 742. raffinose, 738.

trehalose, 720.
Aspergillus oryzae, 692. Assimilation, 532, 533.
Association of Official Agricultural Chemists,
method
for clarifying milk, 447.
xviii
INDEX
method
for determining alcohol precipitate, 520. ash, 495.
Association of Official Agricultural Chemists:
dextrin, 301.
moisture, 16, 18. preparing alumina cream, 223. standard invert sugar, 390.
modification of Sachsse's
Soxhlet's
Tollens's
method for starch, 439. method for reducing sugars, 390 method for galactan, 459, 460.
reports of Referees on sugar, 224, 254.

Astragalose, 729.
Asymmetric carbon atom, 530. Atmospheric pressure, influence on copper reduction, 418. Atomizer for removing foam, 205.
Autoclave, 439, 440.
Autolysis of yeast, 669.
Baeyer's theory of photosynthesis of sugars, 533.

Bacillus
gummosus, 653.
lactis acidi, 583.
levaniformans, 615.
suavolens, 586.
Bacterium gummosum, 653. oxydans, 702.

pediculatum, 584, 652, 653. xylinum, (Sorbose bacterium):
action
upon
alcohols of sugars, 771, 772. 1-arabite, 562.

i-erythrite, 542.
d-galactose, 604. d-glucose, 585.

glycerol, 539.
d-mannite, 617.
perseite, 637. d-sorbite, 624.
sucrose, 654.
Bagasse
(see
under Sugar cane).
Balance, analytical, 39, 162. metric solution, 163.

sugar, 162.
Westphal (Mohr's
specific gravity), 40-42.
Balling's specific gravity table, 29.
Bang's copper bicarbonate method for determining glucose, 434.

Barbituric acid for precipitating furfural, 454.
methylfurfural, 457.
Bardach and
for destroying optical activity of sugars, 304. Barfoed's copper acetate solution, 336, 432. Barium monosaccharate, 680.
Silberstein's
method
process of recovering sucrose, 680.
INDEX
Barium
raffinosate, 739.
xix
Bates's saccharimeter, 139-143.

principle of, 140-142. zero-point error of, 142.
Baumann's
reaction, 370.
Baume hydrometer scale, 48, 49. old and new degrees, 48, 49; Appendix,
Beckmann's apparatus
6.
for determining depression of freezing point, 327, 328. elevation of boiling point, 331, 332.
Benzaldehyde, reaction with sugar alcohols, 766. use in liberating sugars from hydrazones, 348. Benzals of sugar alcohols, 766, 769, 770.
table of formulae, properties, etc., 770.
Benzoyl chloride, reaction with sugars, 369, 370. Benzylphenylhydrazine, 346. Bertrand's method for determining galactose, glucose, invert sugar, kctose and maltose, 426; Appendix, 79.
Bertrand's reaction for xylose, 555, 556.
Betite, 756.
Bial's orcin test for pentoses
and glucuronic
acid, 382.
Bichromate
light filter, 115-117. Biot's polariscope, 84.
Birotation (see Mutarotation)
Bismuth
solution, Nylander's, 338.
Blankit, 221.
Block, Maquenne's, 357. Boiling point of sucrose solutions, 651.

Boiling point of sugar solutions, determination of, by Beckmann's method, 331. application to molecular weight determinations, 332.
Bomb
Bone
calorimeter (see Calorimeter). black, absorption error in sucrose polarization, 220, 221. raffinose polarization, 284, 285.
purification of, 219.
use in decolorizing sugar solutions, 219, 277.
Boot's pycnometer, 38.
Borax and boric
acid, influence
on rotation
of sugar alcohols, 765, 766.
reaction with sugar alcohols, 765. Boring rasp, Keil's, 226. Bornesite, 762.
Brix hydrometer, 44, 45.

specific gravity table, 29; Appendix, 6. Bromine, oxidation of sugars with, 363, 772.
test for aldoses
and
ketoses, 363.
Bromomethy If urf ural, 62 1
p-Bromophenylhydrazine, 347.
test for d-glucuronic acid, 376.
Brown, Morris and Millar's method for determining glucose, fructose and invert sugar, 425; Appendix, 62. Brown, Morris and Millar's theory of diastatic conversion, 686, 687. Browne's diagram of temperature corrections, 258.
XX
INDEX
Browne's formulae for analyzing sugar mixtures, 477-483. method for determining dextrin in honey, 521. commercial glucose in honey, 294.
of
vacuum
drying, 23, 24.
Bryan's results on action of clarifying agents, 224. precipitation of sugars by basic lead, 216, 444. Bryan, Given, and Straughn's method for extracting sugarsj 446.
Butyric fermentation, 583, 584, 652, 715, 716.
Cabinet, for constant temperature polarization, 169. portable polariscope, 170. Calc spar, 80.
Calcium bisaccharate, 677.

monosaccharate, 677.
raffinosate, 739.
Cald well's
trisaccharate, 678. crucible, 415.
Calibration of polariscope tubes, 155, 156. by Landolt's gauge, 155. of sugar flasks, 166-168.
Calories, 313-321. calculation
from formula
centuple, 313.
of sugars, 320, 321.
definitions, 313.
gram-molecular, 318.
large or kilogram, 313. small or gram, 313. determination, 314-318.
table of values for different sugars, 319.
Calorimeter,
bomb, 313-318.
description
and operation, 314, 315.
hydrothermal value, 315, 316.

radiation correction, 316.
Cane sugar
(see
Sucrose)
Caps, Wiley's desiccating, 160, 161. Capillary tube method for determining melting points, 356, 357. Capsules for drying sugar products, 16, 19. Caramel, Ehrlich's colorimetric method for estimating, 467.
preparation, 655. properties, 656.
Caramelane, 656. Caramelene, 656. Carameline, 656. Carbohydrates, 528, 529. formation in nature, 532, 533. Carbon dioxide, method for decomposing saccharates, 250. estimation of in fermentation methods, 460-464. Carbonatation, 646. Care of polariscopic apparatus, 169-171.
Carob beans, preparation
of
d-mannose from, 596.
INDEX
Carr's
xxi
vacuum oven, 22, 23. Cellobiose (see Cellose).

Cellose, 726-728.
octacetate, 727.
preparation from cellulose, 726, 727.

properties, 727, 728. Celloxin, 376. Cellulose, 575.
conversion to cellose, 726, 727. formation by bacteria, 654, 655.

hydrolysis to d-glucose, 580. Cellulosic fermentation, 654.
Centrifugals, laboratory hand, 502. Cerasinose, 560.
Cerealose
(see
Maltose).
Chandler and Ricketts's method of high temperature polarization, 289-291. Cherry-gum, method of hydrolyzing, 548-550. Chips, sugar beet (see under Sugar beet).
Chitin, 752.
hydrolysis, 752. occurrence, 752.
Chitonic acid, 755, 782.

Chitosan, 752. formation from chitin, 752.
hydrolysis, 752. Chitose, 754, 755.
formation from d-glucosamine, 754.

properties, 754. reactions, 755.
oxidation to chitonic acid, 755.

Chocolate, determination of sucrose and lactose Chondroglucose, 631.
in,
280, 281.
Cider vinegar, determination of d-glucose and d-fructose Cinchona bases, use in resolving d, 1-acids, 786, 787.
Citric fermentation, 585, 655, 702.
in,
479.
Citromyces glaber, 655.

Pfefferianus, 702. Clarification, of milk, 447.
of raw-sugars, 204, 207-225. errors in, 207-225.
change in rotation of sugars, 216, 217.

216. precipitation of sugars, 215,
method
volume of precipitate, 209-215. of Herles, 218, 219.
Home,
212-214.
Sachs, 210, 211. Scheibler, 209, 210.
Zamaron, 218.
of solutions for chemical methods, 443, 444. animal substances, 447.
xxii
Clarification of solutions for chemical
INDEX
methods:
plant substances, 443. Clerget methods, 276-278 (see also Clarifying Agents).
Clarifying agents, 207-225.
comparative value of different, 223-225. errors in use of, 209-225.

list of,
alum, 223. alumina cream, 222, 223.

basic lead nitrate, 218, 219.
bone black, 219, 220, 277. dry lead subacetate, 212-215.

hydrosulphites, 221, 222. hypochlorite, 218.
lead acetate, neutral, 207. nitrate, basic, 218, 219.
subacetate, dry, 212-215.

solution, 207, 208.
mercuric nitrate, 447.

sulphites, 278. zinc dust, 278.
Clerget
method
application
of inversion, 264-286. of, to determination of raffmose, 281-286. sugars in presence of sucrose,
279-
281.
factor for, influence of acids, 269.
amino compounds, 270.

concentration, 267, 268. fructose, 270.
modification for impure products, 271-276. invertase method, 274-276. neutral polarization, 271.
urea method, 271-273. use of organic acids, 273, 274. modification of Andrlik and Stanek, 271-273. Herzfeld, 266-268.
Hudson, 275.
Ogilvie, 274.
O'Sullivan and Tompson, 274. Tolman, 269

principle of, 263, 264. reliability of results, 278, 279. Clostridium butyricum, 584.
Coefficient of purity, 494, 495. Colloidal water, 229, 230, 246. Color reactions of sugars, 339-345, 378-386.
d-glucuronic acid, 381-383. ketoses, 378-381.
methylpentoses, 385, 386. pentoses, 381-383. use of spectroscope in studying, 342-345.
INDEX
Color reactions of sugars:
xxiii
with acids, 340, 341.

alkalies, 339.
phenols, 341.
a-naphthol, 378-379.
naphthoresorcin, 381, 383.
orcin, 382.
phloroglucin, 381, 382. resorcin, 380, 381.
Colorimeter, Duboscq's, 464-467. Stammer's, 467-469.
Colorimetric methods for determining caramel, 467. sugars, 464-469.
Combined methods
for analyzing sugar mixtures (see under Mixtures).
Commercial glucose, determination by high temperature polarization, 289-296. method of Browne, 294, 295. Chandler and Ricketts, 28&-291.
Leach, 291-293. estimation in honey, 294-296.
polarizations
of,
293.
mixtures with honey, 296.

process of manufacture, 698. Compensation, of optical activity, molecular, 531. quartz-wedge, 108-112.
double, 110-112. single, 108-110.
Compensator of refractometer, 57, 58. Composition of osazones of sugars, 371.
Compound
sugars, 528.
Concentrating sugar solutions, 448. Concentration, effect on Clerget factor, 267.

rotation of sugars, 174-177.
equations for expressing, 174-177.

viscosity of sugar solutions, 310.
Concentric
field, 93.
half-wave plate, 93.
Conductivity and inverting power of acids, 663.

Coniferin, 571. Contraction of sugar
and water mixtures, 32-34. volume during inversion, 662.
Control-tube, 122-125. Control-wedge, 110-112.
Convallamarin, 599.
sugar, 631.
Convallarin, 599.
Conversion factors, for polariscope Conversion of starch, 685-698.
scales, 145, 1 96-201 o
by
acids, 697, 698.
formation of dextrin, 697.

maltose, 697.
INDEX
Conversion of starch by acids:
formation of reversion products, 697.
technical processes, 698.
by enzymes, 685-696.
malt diastase, 685-692.
influence of acids, alkalies, etc., 691, 692. temperature, 690.
restriction of, 690, 691.
steps of process, 686.
theory of
Brown and coworkers, 686, 687. Maquenne and Roux, 687-689.

of,
pancreatin, 693-696. activation
694.
converting power of highly active, 696.

influence of acids
and
alkalies, 694.
concentration of starch, 695.
temperature, 695, 696.

ptyalin, 693.
takadiastase, 692, 693.
Convolvulin, 567, 569.
Coomb's drip sampler,

Copper, ferrocyanide
10, 11.
test,
392-394.
hydrobromic methods of determining, 403-417.
of Wiley, 393. acid test, 410.
method method
of Ross, 393, 394.
by
electrolysis,
406-410.
from
nitric acid, 407.
sulphuric acid, 406, 407.

sulphuric and nitric acids, 407. tartrate solution, 409, 410. by reduction of cuprous oxide in hydrogen, 403-405.
by
titration, 410-415.
volumetric cyanide method, 415. iodide method, 411-414.

modification of Kendall, 412, 413.
Low, 411, 412.

Peters, 413, 414.
permanganate method, 410, 411. thiocyanate method, 414, 415.
by weighing
as cupric acid, 415, 416. cuprous oxide, 416.
comparison of methods, 416, 417. Copper-reducing power of sugars, 421-423. Copper reduction, factors influencing, 417-419.
atmospheric pressure, 418, 419. dilution of solutions, 417, 418. purity of reagents, 417.
surface area of solution, 419. temperature, 418, 419.
INDEX
Copper reduction, factors influencing:
time of boiling, 417, 418. Copper-reduction methods for determining sugars, 388-435.
xxv
method
of Allihn, 403.
Bang, 434, 435.

Barfoed, 432.
Bertrand, 426. Brown, Morris and Millar, 425. Defren, 425, 426.
Fehling, 389. Herzfeld, 428. Kendall, 435.
Kjeldahl and
Meissl, 423.
Woy, 424, 425. Koch and Ruhsam, 420.
Meissl and Hiller, 430, 431. Meissl and Wein, 428-430.
Munson and Walker,

Ost, 433, 434.
426, 432.
Pavy, 395-397.
Pfliiger, 419, 420.
Reischauer and Kruis, 398, 399.

Soldaini, 432.
Soxhlet, 389-391, 424.

Violette, 393-395. Wein, 423.
Corrections, temperature (see under Temperature).
Cottonseed meal, preparation of raffinose from, 733, 734. Cover-glasses for polariscope tubes, 156.
Creydt's formula for estimating raffinose, 282. Crystal content of raw sugars, determination of, 498-506. method of Herzfeld and Zimmermann, 503-506.
Koydl, 501, 502. Payen, 499. Scheibler, 499-501.

Crystalline forms of sodium
ammonium
tartrate, 785.
Cubic
sucrose, 647, 648. centimeter, 27, 28. metric, 28.
Mohr,
28.
reputed, 28.
Cupric oxide, determination of copper by weighing, 415, 416. Cuprous oxide, contamination of, 416, 417. determination of copper by weighing, 416.
method
of filtering, 404.
reduction in hydrogen, 403-406. Cyanhydrine reaction of sugars, 365, 366. Cyanide metnod for determining unreduced copper, 415.
Cy closes,
Cyclamose, 560. 755-763.
xxvi
INDEX
determining specific gravity, 45.
Cylinders, for filtering sugar solutions, 168.
Cytase, 686.
and d, 1-, meaning Dambonite, 762.

d-
of prefix, 532.
Dambose
(see i-Inosite).
(see Clarification
Decolorization of sugar solutions Decoses, 642.

Decrolin, 70.
and Clarifying
agents).
Defecation
Defren's
(see Clarification
method
for determining glucose, lactose acids, 781.
and Clarifying agents). and maltose, 425, 426; Appendix,
63.
Dehydration of hexose dibasic
Dehydromucic
acid, 781, 782.
formation of furfural from, 782.

reaction for hexose dibasic acids, 781. test of Tollens and Yoder for, 781.
Deleading sugar solutions, 276, 277. Depression of freezing point (see Freezing point). Desiccating caps, Wiley's, 160, 161. Destruction of optical activity of reducing sugars, 302-306.
by
alkalies, 302.
method
of
Bardach and Silberstein, 304, 305. Dubrunfaut, 302, 303.

Jolles, 304.
by
Lobry de Bruyn and van Ekenstein, 303. and hydrogen peroxide, 305, 306. method of Pellet and Lemeland, 305. by alkalies and mercuric cyanide, 306. method of Wiley, 306.
alkalies
Deterioration of raw sugars, 14. Dextran, 578, 584, 653, 654.

formation by bacteria, 584, 653, 654. influence on polarization of sugar products, 654.
properties, 584.
Dextrin, 577, 686-691.
commercial, composition
of,
510.
of analysis, 508-510. process of manufacture, 577. viscosity of solutions, 508, 510.
methods
determination by Fehling's solution, 442.

fermentation, 301, 302. precipitation with alcohol, 490. Wiley's method, 490.
in fruit products, 301, 302.
honey, 521-523. presence of glucose and maltose, 486-488, 490-492. formation during conversion of starch, 686-691. plant-, 578.
INDEX
Dextrin, researches of Dextrinase, 686.
xxvii
Brown and
Millar on, 687.
Dextrinomaltase, 686.
a-
and /3-Dextro-metasaccharin, 587.

(see
Dextrose
d-Glucose).
Dhurrin, 573.
Diarabinose, 643. Diastase, 683-685.
action on starch (see under Conversion).
formation during germination of barley, 683. method for determining starch, 440-442.
occurrence, 683. preparation from malt, 685.
properties, 685.
Diastatic-power, methods of determining, 511-515. method of Lintner for diastases, 513.
method
of
malt and malt extracts, 511-513. Sherman, Kendall and Clark, 513-515. Sykes and Mitchell, 513.
Wohlgemuth, 515.
Digestion methods for polarizing sugar beets
Digitalin, 570. Digitalose, 570.
(see
Sugar beets).
Digitonin, 599. Digitoxin, 544.

Digitoxose, 543, 544.
Dihexose saccharides, 645-730.

Dilution, double, 209, 210.
effect
on copper-reduction, 417, 418.

specific gravity, 35.
of solutions in determining refractive index, 66-69.
Dimethyldioses, 537.
Dimethylglycolose, 537.
Dimethylketol, 537. Dimethyltetroses, 543, 544.

Dioses, 535.
Dioxyacetone, 538, 539. Dipentose saccharides, 643.

Diphenylhydrazine, 346. Disaccharides, 643-730.
variability in reducing
power
of,
402.
Dissociation and inverting power of acids, 663-666. salts, 666-668.
Double
dilution,
method
of Scheibler, 209, 210.
Wiley and Ewell, 253.

Double-field, 89-94.
Double hydrazides, 782. Double lactones, 780. Double quartz plate, Soleil's, 86-88. Double refraction, 80.
xxviii
Double-wedge system, 110-112.
INDEX
Dry lead subacetate, 212-214. Dry substance (see Total solids).

Dubois's method of determining lactose and sucrose, 280, 281. Duboscq's colorimeter, 464r-466.
saccharimeter, 132, 135.
Dubrunfaut's method of destroying optical activity of sugars, 302.

Dulcite, calorific value, 319. dibenzal, 770.
formation by reducing d-galactose, 606.

occurrence, 606. oxidation to d, 1-galactose, 607.
Dutch standard,
properties, 768. 498.
method for estimating caramel, 467. Einhorn's fermentation saccharometer, 462, 463.
Ehrlich's colorimetric
Electric lamp,
Schmidt and Haensch, 153.
stereopticon, 152. Electrolytic apparatus of Leach, 407-409. determination of copper (see under Copper). Elementary composition of osazones, 371.
Eliett
and
Tollens's
method
for determining methylpentoses
and methylpentosans,
456-458; Appendix, 89. Elution process of Scheibler, 678.
Emulsin, action upon glucosides, amygdalin, 572.

dhurrin, 573. a- and /3-glucosides, 591.
prulaurasin, 572. salicin, 571.
sambunigrin, 572.
saccharides, cellose, 727.
galactosido-galactose, 728. gentiobiose, 726. glucosido-galactose, 728.
isomaltose, 705.
raffinose, 737.
occurrence, 572. synthetic action Engler's viscosimeter, 308.
upon
d-glucose, 704, 705.
Enzymes, acting upon glucosides,

emulsin, 571-573, 591. indimulsin, 571.
maltase, 591. myrosin, 573. tannase, 573.

acting
upon
saccharides,
amylases, 683-696.
cytase, 686. diastase, 683, 685-692.
INDEX
Enzymes, acting upon saccharides:
dextrinase, 686.
xxix
emulsin, 726-728, 737.

inulase, 615.
invertase, 651, 668-676, 737, 743, 748.

lactase, 713, 714.
maltase (maltoglucase), 686, 701, 702.

melibiase, 723.
pancreatin, 693-696. ptyalin, 693.

takadiastase, 692, 693. trehalase, 720.
zymase, 582
Enzymic
synthesis, 704, 705 d-Erythrite, 542, 767.
1-Erythrite, 542, 767.
i-Erythrite (mesoerythrite), 541, 767. calorific value, 319.

dibenzal, 770. occurrence, 541.
oxidation to d, 1-erythrose, 541.
Erythrodextrin, 577, 686.

d-Erythrose, 540, 541. 1-Erythrose, 541. d, 1-Erythrose, 541, 542.
d-Erythrulose, 542, 543. d, 1-Erythrulose, 543.
Ester fermentation, 586.

Ether, atomizer, 205. use in purification of sirups, 550. Ethylphenylhydrazine, 346.
Evaporation of moisture from raw sugars, 8, 9. sugar solutions in vacuum, 549-550.

Extraction, determination of, 496. Extraction of sugars, alcoholic, 233, 446.
method
of Bryan,
Given and Straughn, 446.
Scheibler, 233-235.
aqueous, with cold water, 445 with hot water (Zamaron), 235-238."
Expression of juice, 227-230.
errors of
method, 229.
for, 227,
hydraulic press
228.
Fehling's copper solution, 335, 389-444. composition, 335, 389. factors influencing results (see under
Copper reduction). gravimetric methods employing, 399-443. products obtained by action on sugars, 335, 336.
reducing action of sucrose on, 427. use in determining dextrin, 442.
xxx
Fehling's copper solution:
INDEX
use in determining glycogen, 443. starch, 438-442.
sucrose, 436-438.
volume reduced by different sugars, 391. volumetric methods employing, 389-399.

Fermentation, alcoholic, 581, 582, 651.
butyric, 583, 584, 652. cellulosic, 654, 655.
citric,
585, 655.
ester, 586.
gluconic acid, 585.

lactic, 583, 652.
mannitic, 653, 654.

oxalic, 585.
viscous, 584, 652.
Fermentation flask, 300. Fermentation methods for determining sugars, 299-302, 460-464. by Einhorn's saccharometer, 462, 463.
Lohnstein's saccharometer, 463, 464. weighing carbon dioxide, 461, 462.
resolving racemic mixtures, 787.
Fermentation of raw sugars, 14. Fermentations of sugars: 1-arabinose, 550, 551.

d-fructose, 619.
d-galactose, 604. gentianose, 743.
d-glucose, 581-586. isomaltose, 707.

lactose, 713-716.
maltose, 701-703.
mannatrisaccharide, 745.
d-mannononose, 641. d-mannose, 596, 597.

melibiose, 723. raffinose, 738.
rhamnose, 565.
d-sorbose, 625. stachyose, 748.
sucrose, 651-655. trehalose, 720.
1-xylose, 555.
Ferrocyanide test for copper, 392, 393. Fiber, determination in bagasse, 248.
sugar beets, 228, 229.
Field of vision in polariscopes
:
concentric, 93. double, 89-94.

fringed, 100.
quadruple, 97, 98.

triple, 97, 98.
INDEX
Fillmass, 646 (see Massecuite)
.
xxxi
Filter-press cake, polarization of, 249-251. in absence of saccharate, 249, 250. presence of saccharate, 250, 251.
acetic acid method, 250. ammonium nitrate method, 251.
carbon dioxide method, 250. zinc nitrate method, 251.

Filter-tube, Knorr's, 393. Wiley's, 393.
Filtration of sugar solutions, 205. Fischer's hydrazone and osazone reaction of sugars, 345-362. method of oxidizing alcohols to sugars, 770, 771.
reducing lactones to sugars, 776. synthesis of d-fructose, 355, 622, 623. d-galactose, 602.
d-glucose, 580. isomaltose, 705.
d-mannose, 596. methyl glucosides, 590.

Flasks, calibration of, 166-168. for fermentation, 300.
polariscopic analysis, 163-168. solution by weight, 164.
volumetric use, 165. specifications for, 166.
Formaldehyde, reaction with sugar alcohols, 766. use in liberating sugars from hydrazones, 348. Formals of sugar alcohols, 766. Formation of carbohydrates in nature, 532-534.
Formose, 629, 630. /3-Formose, 630. Frangulin, 563.
Fraunhofer's
lines, 343, 384.
Freezing point of sugar solutions, 325-331. application to determining molecular weights of sugars, 329-331.
rate of inversion, 331.
molecular depression of, 329. Raoult's method for determining depression

relation to
of,
327-331.
by Beckmann's apparatus, 328.

vapor and osmotic pressure, 326, 327.
French sugar
scale, 112, 113.
value in circular and Ventzke degrees, 145.

Fric's saccharimeter, 138, 139.
illuminating device Fringes, interference, 100.
of,
137.
d-Fructose, 612-622.

resorcin, 381, 384.
action of alkalies on, 339, 340.
xxxii
d-Fructose, calorific value, 319.
color reactions, 378, 619.
INDEX
*
Seliwanoff 's resorcin test, 380.
decomposition into oxymethylfurfural, 620. determination, as methylphenylosazone, 470. by copper reduction methods,
424,
425
(see
under
Copper reduction), by polarization at high temperature, 296-298. Sieben's method, 470, 471.
in cider vinegar, 479.
presence of 1-arabinose, 482. d-galactose, 481.

d-glucose, 477-479.
d-glucose and d-galactose, 484. d-glucose and sucrose, 485, 489. of moisture in fructose products, 20.
effect of
temperature on polarization, 179, 297, 478. fermentation, 619. action of different yeasts, 714. formation by hydrolysis from gentianose, 743.
inulin, 618.
lupeose, 749. melezitose, 742.

raffinose, 736, 737.
secalose, 746.
stachyose, 748. sucrose, 617, 660.

turanose, 725. verbascose, 750.
influence on Clerget factor, 270.
methylphenylosazone reaction, 621, 622.

mutarotation, 187, 618.
normal weight, 197.

occurrence, 612-616.
osazone, 354, 622 (see d-Glucose-osazone). influence of lactose, maltose and sucrose on formation
353.
of,
352,
precipitation
oxidation with bromine, 363, 619. by basic lead salts, 216.
preparation from inulin, 618.

sucrose, 617, 618.
properties; 618.
protective action on invertase, 675, 676. reaction with hydrobromic acid, 621.
reducing ratio to glucose, 391, 421. reducing reactions, 621. reduction to d-mannite and d-sorbite, 619.
specific rotation, 173-192, 618.
influence of acids on, 185, 186. alcohol on, 181, 182.
INDEX
d-Fructose, specific rotation, influence of lead subacetate on, 185, 217. urea on, 272.
synthesis from d-glucose
xxxiii
and d-mannose by action

616.
reduction
of alkalies, 303. of osones,
355,
d-mannite, 617.
tests,
619-622.
value of Ventzke degree, 200, 201. yield of levulinic acid from, 373. 1-Fructose, 622. d, 1-Fructose, 622, 623.
synthesis from acrolein dibromide, 623. Fruit products, determination of alcohol precipitate in, 520. Fuconic acid, 566-568.
Fucosan, 565. determination, 457; Appendix, 89

Fucose, 565, 566.
(see also
under Methylpentosans).
determination, 456, 457; Appendix, 89 mutarotation, 187, 566.

occurrence, 565. preparation, 565, 566. properties, 566.
(see also
under Methylpentoses).
racemic combination with rhodeose, 568.

specific rotation, 566.
tests, 566.
Funnels for
yield of methylfurfural from, 377. filtering sugar solutions, 168.
transferring sugars, 203. Furaloid, 453.

Furfural, apparatus for distilling, 450, 451.
determination, 449-455. formation from d-glucuronic acid, 375, 376.

oxycellulose, 376, 377.
pentoses and pentosans, 374.
method
for determining pentoses
and pentosans, 449-455.
phenylhydrazone, 375.
phloroglucide, 375, 451. factors for converting to pentoses and pentosans, 452. precipitation with ammonia, 449.
barbituric acid, 454.
phenylhydrazine, 449.
phloroglucin, 451. reaction for pentoses and pentosans, 374, 375. limitations
yield Furfuran, 755.
of,
375-377.
from pentoses, 374.
Furfuroids, 453.
'
xxxiv
Galactan, 599, 600.
determination, 459, 460.
INDEX
Galactoaraban, 599, 600. Galactoarabinose, 644.
Galactomannan, 600.
Galacto-metasaccharins, 604. d-Galactonic acid, conversion to lactone, 774.
d-talonic acid, 611, 775.
oxidation to d-lyxose, 557. lactone of, rotation, 774.

1-Galactonic acid, 606.
d, 1-Galactonic acid, 608.
resolution of, 608, 787.
d-Galactose, 598-606.

resorcin, 381.
action of alkalies on, 603, 604, 625, 626. calorific value, 319.
conversion to a- and /3-galaheptose, 636. d-tagatose and 1-sorbose, 626.

d-talose, 611, 777.
determination by copper reduction, 426.
mucic acid method, 459.

in presence of d-fructose, 481.
d-glucose, 480. d-fructose and d-glucose, 484. effect of temperature on polarization, 179, 480. fermentation, 604.
action of different yeasts, 714.
formation by hydrolysis from galactans, 599, 600.

lactose, 602, 713.
lactosinose, 746. lupeose, 749.
pectins, 601.
raffinose, 736.
rhamninose, 732.
stachyose, 748.
verbascose, 750.
hydrazones, 605.
modifications, 192, 603. mucic acid reaction, 459, 460, 604, 605. mutarotation, 187, 603.
occurrence, 599-602. osazone, 605.
oxidation with bromine, 363, 606. preparation from agar-agar, 603.

milk-sugar, 602, 603.
properties, 603.
reducing ratio to glucose, 421.
INDEX
d-Galactose, reduction to dulcite, 606.
xxxv
synthesis, 602.
value of Ventzke degree, 200, 201.

variability in reducing power, 400. yield of levulinic acid from, 373. mucic acid from, 459.
1-Galactose, 606, 607.

d, 1-Galactose, 607, 608.
Galactosido-galactose, 728. Galactosido-glucoheptose, 730.
Galactoxylan, 600. Galaheptite, 768. a-Galaheptonic acid lactone, rotation a-Galaheptose, 636.
/3-Galaheptose, 636. Galaoctite, 768.
of,
774.
a-Galaoctonic acid lactone, rotation

a-Galaoctose, 639, 640. Galtose, 628, 629.
of,
774.
Gas lamps, 152. Gas pressure, relation
to osmotic pressure, 323.
Gauge
for calibrating polariscope tubes, 155.
Gaultherin, 571.
to diarabinose, 643. Geerlig's refractometer table, 65; Appendix, 22. researches upon inverting power of invert sugar theory of melassigenic action, 650, 651.
Gedda gum, hydrolysis
and
salts, 667, 668.
Gentianose, 726, 742-744. action of enzymes on, 743.

configuration, 743. fermentation, 743.
hydrolysis, 726, 743. occurrence, 742.
preparation, 742.
properties, 743. Gentiobiose, 726, 743.
formation from gentianose, 726, 743.
German
preparation and properties, 726. or Ventzke sugar scale, 113-115 (see also under Scales).
82.
Glan prism,
Glucase, 591, 683, 701, 702. Glucoapiose, 643, 644.
formation from apiin, 643, 644.

hydrolysis to apiose a-Glucodecite, 642.
and
d-glucose, 644.
a-Glucodecose, 642.
Glucogalactan, 599.
Glucoheptite, 768
monobenzal, 770.
xxxvi
INDEX
a- and /3-Glucoheptonic acids, 633, 634. rotation of lactones, 774. a-Glucoheptose, 633.
action of different yeasts upon, 714. conversion to a-glucooctose, 638.
0-Glucoheptose, 634. d-Gluconic acid, conversion to d-mannonic acid, 775.

fermentation, 585. formation from d-glucose
by oxidation by
bacteria, 585.
with bromine, 590.

lactone, rotation of, 590, 774. oxidation to d-arabinose, 545.
1-Gluconic acid, synthesis from 1-arabinose, 592. a-Glucononite, 641, 768.
a-Gluconononic acid, 641.

a-Glucononose, 640, 641. conversion to a-glucodecose, 642.
a-Glucooctite, 638, 768. a- and j8-Glucooctonic acids, 638. rotations of lactones, 774. a-Glucooctose, 638.
action of different yeasts upon, 714. conversion to a-glucononose, 641.

/3-Glucooctose, 638. Gluco-proteids, 579.
d-Glucosamine, 751-754.
chloride, 753.
formation from chitin, 752.

mucins, 752.
occurrence, 751.
preparation from lobster

properties, 753.
shells, 752.
synthesis from d-arabinose, 754. tests, 753.
d-Glucose, 570-591.

resorcin, 381.
action of alkalies on, 339, 340, 586, 587. calorific value, 319.
commercial
(see
Commercial
glucose),
conversion to d-arabinose, 365.

d-glucoheptose, 365.
dehydration of, 25. determination by copper reduction, 389-435
(see
under Copper reduction),
in cider vinegars, 479. in presence of d-fructose, 477-479.
d-galactose, 480. d-fructose and d-galactose, 484, d-fructose and sucrose, 485, 489.
maltose and dextrin, 486, 490.
INDEX
d-Glucose, fermentations
of, 581-586. formation by hydrolysis from:
xxxvii
cellose, 727.
cellulose, 580.
gentianose, 743. gentiobiose, 726.

glucosides, 563, 570-573.
glycogen, 579.
isomaltose, 705. lactose, 713.
maltose, 701.
melezitose, 725, 742. melibiose, 723.
raffinose, 736.
stachyose, 748. starch, 580.

sucrose, 581. trehalose, 720.
turanose, 725. verbascose, 750.
hydrazones, 589.
influence of sucrose on reducing power, 427. urea on polarization, 272.
manufacture
of,
698.
modifications, 192, 581.
mutarotation, 187-193, 581. normal weight, 197-199.

occurrence, 570-580. osazone, 348-354, 589, 590. influence of lactose, maltose, raffinose and sucrose on formation of, 351, 352.
oxidation to d-gluconic acid, 585, 590. with bromine, 363.

precipitation
by
basic lead salts, 216.
preparation from cellulose, 580.
honey, 580.
starch, 580. sucrose, 581.
properties, 581.
reactions, 362-370, 587-591. reduction to d-sorbite, 590.
saccharic acid test, 587, 588.

synthesis, 580.
587-590. value of Ventzke degree, 200, 201. variability in reducing power, 400.
tests,
yield of levulinic acid from, 373. 1-Glucose, 592, 593.
xxxviii
d, 1-Glucose, 593.
INDEX
Glucose ratio, 496. Glucose reduction equivalents of sugars, 421, 476. Glucosides, glucose-yielding, 570-573.
preparation from plant substances, 574.
rhamnose-yielding, 563, 564. synthetic, 590, 591.
Glucosido-galactose, 728. Glucosido-glucoheptose, 730. Glucosuria, 571, 578.
Glucotannin, 573. d-Glucuronic acid, color reactions with naphthoresorcin, 383.

orcin, 382.
phloroglucin, 382. conversion to 1-xylose, 375.
formation from d-saccharic
ajcid,
608, 783.
occurrence in urine, 375, 783. production of furfural from, 375, 783. reaction with p-bromophenylhydrazine, 376.
Glutose, 629. occurrence in cane molasses, 629. Glyceric aldehyde (see d, 1-Glycerose) Glycerol, 538, 767.
.
monobenzal, 770. oxidation by Bacterium xylinum, 539.

to dioxyacetone, 539. d, 1-glycerose, 538.
d, 1-Glycerose, 538.
antipodal forms Glycogen, 578, 579.

of,
530.
determination by Fehling's solution, 443. occurrence, 578.

preparation, 578, 579. properties, 579.
vegetable-, 578. Glycol, 535, 767. Glycolaldehyde, 535.
Glycolose, 535.
Gooch
crucible, 404, 415.
Gossypose (see Raffinose). Graduation of hydrometers, 43-49.

saccharimeter scales, 117-119.
Gram-molecular calories
(see Calories).
Graminin, 615. Grape sugar (see d-Glucose). Gravimetric methods for determining sugars, 399-445. d-Gulonic acid, conversion to d-idonic acid, 610.
formation from d-saccharic acid, 608. oxidation to d-xylose, 552.
INDEX
d-Gulonic acid, rotation of lactone, 609, 774. 1-Gulonic acid, conversion to 1-idonic acid, 775. formation from 1-xylose, 609.
d, 1-Gulonic acid, 610.
xxxix
hemihedry
of lactone crystals, 610, 786.
d-Gulose, 608, 609. conversion to d-idose, 610, 777.

1-Gulose, 609, 610. action of different yeasts upon, 714. d, 1-Gulose, 610.
Gums,
solution of in digesting sugar beets, 245.
Half-shadow, angle, 89-92, 94-96.

polarimeters, 89-98, 101-106.
saccharimeters, 132-145. Half-wave plate, Laurent's, 91-93.
Hayduck's nutritive
salt solution, 299.
Hay wood's
Heat
of
modification for determining methylpentoses, 458, 459.

(see Calories).
combustion
Hederose, 631.
Helianthenin, 615.
Hemicelluloses, 439, 441, 534, 546, 553, 575, 593, 599. crystals, of d, 1-gulonic lactone, 610, 786.
Hemihedral
sodium ammonium
Heptoses, 633-637.
Herles's basic lead nitrate
raffinose
d, 1-tartrate, 785.
surfaces of sucrose crystals, 647.
method
of clarification, 218, 219.
formula for variations in temperature, 283, 284. Herzfeld's method for determining acidity and alkalinity, 496, 497.
invert sugar in raw sugars, 428; Appendix, 81. raffinose, 282, 283.
method
of alcoholic digestion
and extraction, 247, 248.
hot-water digestion, 244.

preparing maltose, 699. modification of Clerget's method, 266-268.
Herzfeld and Zimmermann's method for determining crystal content, 503-506.
Hesperidin, 563.
Hexose groups,
levulinic acid reaction for, 372-374.
Hexose-heptose saccharides, 730. Hexoses, 570-631. " High-polarizing" sugar, 658, 659. Hinks's oil and gas lamps, 151.
Honey, 579, 580, 616.

detection of artificial invert sugar
in,
620.
commercial glucose in, 523. determination of commerical glucose in, 294-296.

dextrin
in,
521-523.
dextrorotation at 87 C. after inversion, 293, 294. occurrence of fructose, glucose and sucrose in, 616.
polarization of varieties, 294.
xl
INDEX
of varieties containing commercial glucose, 296. preparation of d-glucose from, 580.
Honey, polarization
table of composition, 522.
Honey-dew, 522.
occurrence of melezitose
in,
740.
516, 517. Hiibener's refractometer table, 74; Appendix, 24. Hudson's constant temperature water bath, 160.
Home's method of dry defecation, 212-215. Hortvet's method for measuring lead precipitate,
equation for inversion of sucrose, 672. modification of Clerget's method, 275.

researches
upon
invertase, 669-676.
lactose, 710, 711.
rotation of lactones, 774, 775.
Humus
"
substances, 340.
polarization," 125, 126.
Hundred
Hydraulic laboratory press, 227, 228. Hydrazides of dibasic acids, 782.
monobasic
acids, 777.
Hydrazines, optically active, for resolving d, 1-sugars, 361, 362, 551.

substituted, 346, 347.
Hyclrazone reaction of sugars, 345, 346.
Hydrazones, analysis of, 370, 371. decomposition of, with benzaldehyde, 348. formaldehyde, 348.
hydrochloric acid, 347. determination of sugars from weight of, 469, 470.
identification, 356-360, 370. melting point of (see Melting points),
optical activity of, 360.
separation of sugars from, 347, 348. table of melting points and properties, Appendix, 90 Hydrobromic acid, relative inverting power of, 663.
test for
unreduced copper, 410.
Hydrochloric acid, decomposition products of sugars with, 340, 372-378.

relative inverting power of, 663. Hydrocyanic acid, action upon reducing sugars, 365, 366. Hydrogen, reduction of cuprous oxide in, 403-406.
Hydrolysis of sugar-yielding substances, Tollens's method, 548-550. Hydrometers, 42-49.

according to Baume, 48. Brix, 44-46.
Langen, 47, 48.

Volquartz, 46, 47.
Vosatka, 47. standardization of, 43, 44.
"sweet-water," 47, 48. Hydrosulphites as decolorizing agents, 221, 222. errors due to use of, 222.
Hydrothermal value, 315, 316.
INDEX
Hydroxylamine, action upon reducing sugars, 364, 365. standard solution for titrating copper, 434r-435. Hypochlorite as a decolorizing agent, 218.
i-,
xli
meaning
of prefix, 532.
d-Idite, 610, 767.

1-Idite, 611, 768.
d-Idonic acid, 610.

1-Idonic acid, 611.
conversion to 1-gulonic acid, 775.

d-Idose, 610. 1-Idose, 611.
Illumination of polariscopes, 146-153

refractometers, 58 Imbibition water, 229-230, 246.
(see
under Lamps).
Immersion refractometer, 70-75. adjustment
of,
72-74.
for, 74;
Hiibener's table
Appendix, 24.
principle of, 71, 72.
tempering bath
Imperial
for, 74, 75.

1.
sucrose specific gravity table, 30; Appendix, Incrusting substances, 553, 575. Index of refraction (see Refractive index).
Indican, 571. Inosinic acid, 558.
Inosites, 757-763.
German Commission,
isomeric forms, 757, 758.

d-Inosite, 758, 759.
preparation from pinite, 758.

properties
1-Inosite, 759.
and
tests, 758.
preparation from quebrachite, 759.

properties d, 1-Inosite, 760.
i-Inosite,
and
tests, 759.
760-763.
formation from bornesite, 762.
dambonite, 762.
phytin, 762, 763.
occurrence, 760.
preparation from meat, 760.
walnut leaves, 761

properties and tests, 761, 762. Interference fringes, 100.
International Commission,
Inulase, 615. Inulenin, 615.
Inulin, 613-615.
method
for determining moisture, 16.
rules for polarizing sugars, 201, 202.
hydrolysis to d-fructose, 615, 618.
xlii
Inulin, occurrence, 613.
INDEX
'
preparation
of,
613, 614.
preparation of d-fructose from, 618.

properties, 614.
sphere-crystals, 614.
Inversion of sucrose, 263-279, 659-676.
by
acids, 263-279, 659-666.
invertase, 668-676 (see Invertase).

salts,
666-668.
Clerget's
method (see Clerget). early investigations, 659, 660. hypothesis of Arrhenius, 664.
law
of, 263, 264. rate of, 660-667, 671-674.
determination by freezing point method, 331.

polariscope, 661, 662. influence of concentration, 665. organic substances, 666.
salts, 665.
temperature, 664.
urea, 272, 273. Wilhelmy's law, 660, 661. relative power of acids in, 663.
Invert sugar,
artificial,
620.
color reactions for, 620.
decolorization of solutions, 277, 278.
determination by copper reduction, 423-432 (see Copper-reduction methods) determination by polarization at high temperature, 287-289.
.
in presence of d-glucose
influence
on inverting power normal weight, 197.
and d-fructose, 478. sucrose, 428-432. of salts, 667, 668.
preparation of standard solution, 390. reducing ratio to glucose, 391, 421. specific rotation, 174-192.
influence of acids on, 185, 186. alcohol on, 181, 182.
concentration on, 177. lead subacetate on, 185.
temperature on, 179, 180. urea on, 272.
temperature of optical inactivity, 287, 288. value of Ventzke degree, 200, 201.
variability in reducing power, 400 Invertase, action upon gentianose, 743, 744. raffinose, 737, 738.
stachyose, 748.
sucrose, 668-676.
verbascose, 750.
INDEX
Invert ase, conditions affecting activity
of,
670-676.
alcohol, 674, 675. alkalies, 671.
influence of acids, 671.
concentration of invertase, 673. sucrose, 673, 674.
temperature, 674.
occurrence, 668, 669. preparation, 669, 670.
properties, 670.
protective action of fructose and sucrose on, 675, 676. researches of Hudson, 669-676.
use in Clerget's method, 274, 275.
Inverting action of honey, 13, 616. power of acids, 662-666.

salts,
666-668.
Iodide methods for determining copper, 411-414 (see under Copper), lonization and inverting power of acids, 663-666.
salts,
666-668.
Irisin, 615.
Isatin reaction for
dehydromucic
acid, 781.
Isodulcite (see Rhamnose). Isolactose, 718.
Isomaltose, 705-707. action of emulsin on, 705. preparation, 705.

properties, 707. theories regarding formation, 706, 707. tests, 707.
Isorhamnonic
acid, 568.
rotation of lactone, 568, 774.
Isorhamnose, 568.
Isorhodeose, 569. Isosaccharic acid, 755, 782. Isosaccharin, 713.
Isosaccharinic acid, 712, 713. Isotonic sugar solutions, 326, 327. Isotrehalose, 728, 729.
Ivory nuts, preparation of d-mannose from, 595

Jellet-Cornu prism, 89-91, 133.
Jellet half-shadow polariscope, 89.
Jolles's
method
for destroying optical activity of sugars, 304. determining methylpentoses, 457.
pentoses, 454, 455.

Juice, bottle for weighing, 24. composition of from different cane-mills, 232.
determination of moisture
in,
18-25.
distribution in tissues of sugar cane, 231.
hydraulic press for expressing, 227-229.
xliv
Juice,
INDEX
methods
normal,
of polarizing, 205, 206.
497.
sampling, 10, 11.
Kahlenberg, Davis and Fowler's researches on inversion, 331, 666, 667.

Kefir, 714.
ferment of, 715, 718. Keil's boring rasp, 226.

Kendall's copper-salicylate
iodide
method for determining sugars, 435. for determining copper, 412, 413. Ketoses, characteristic group of, 527.
method
reactions, 340, 341, 354, 363, 364, 378-381. color reactions of (see Color reactions).
conversion of aldoses into, 355.

distinguishing from aldoses (see Aldoses). methylphenylhydrazine reaction, 354.
oxidation with bromine, 363. nitric acid, 364.
Ketoheptoses, 637. Ketohexoses, 612-630.
Ketopentoses, 560-562.
Ketotetroses, 542, 543. Ketotrioses, 538, 539.
Kjeldahl and Woy's method for determining d-fructose, d-glucose, invert sugar,
lactose
and maltose, 424, 425; Appendix,
44.
Knapp's mercury
Knorr's
filter
solution, 338, 435. mercuric cyanide method for determining sugars, 435.
tube, 393.
Koch and Ruhsam's method
for determining glucose, 420; Appendix, 35. Koydl's method for determining crystal content, 501, 502. Krober's factors for calculating pentoses and pentosans, 452. table for calculating pentoses and pentosans, Appendix, 83. Kriiger's automatic pipette, 240, 241.
cold water digestion process, 240-242.
Kumiss, 714.
1-,
meaning
of prefix, 532.
Lactase, 713-715. preparation, 715. Lactic aldehyde, 535, 536.
fermentation, 583, 652, 715. Lactobionic acid, 644, 712.
Lactones of dibasic sugar acids, 779, 780. double lactones, 780.

reduction
lactone acids, 779, 780. of, 782, 783.
monobasic sugar
acids, 773-776.
reduction to sugars, 776.

specific rotation of, 774.
relation of, to configuration, 774, 775.
INDEX
Lactones of monobasic sugar acids:
transformation into acids, 773.
Lactose, 708-718.
xlv

resorcin, 381.
action of acids on, 713.

alkalies on, 712, 713.
enzymes
on, 714.
calorific values, 319.
compounds, 716, 717.

acetates, 716, 717. lactosates, 717.
nitrates, 716.
osazone, 716. configuration, 717. conversion to galactoarabinose, 644. dehydration, 25.
determination by copper-reduction, 424-426 (see Copper reduction methods). polariscopic methods, 252-255. in milk, 252-255. milk chocolate, 280, 281.
milk sugar, 255.

fermentations, 713-716. action of different yeasts, 714. formation of isosaccharin from, 712, 713.
mucic acid from, 712.

hydrolysis
influence
of,
713.
of fructose
on osazone formation
and
glucose, 352.
modifications, 709-711.
inutarotation, 187, 709-711. normal weight, 197, 198.
occurrence, 708. oxidation products, 712.
preparation, 708, 709. preparation of d-galactose from, 602, 603. properties, 709-711.
reactions, 711-713.
reducing ratio to glucose, 391, 422. reduction products, 711. specific rotation, 173-192, 709-711.
tests, 717.
value of Ventzke degree, 200, 201
variability in reducing power, 402. Lactosinose (Lactosin), 745, 746.
occurrence, 745. preparation, 745. properties, 745, 746.
Lamps
for polariscopes, 146-153.
sodium
light,
147-151.
Landolt's, 148.
INDEX
Lamps
for polariscopes
:
sodium
light,
Pribram's, 148.
Zeiss's, 148, 149.
white
light,
151-153.
acetylene, 152. alcohol, 152.

electric, 152, 153.
gas, 152.
oil,
151.
Landolt's concentration formula for rotation of sucrose, 118, 177.
gauge for calibrating tubes, 155, 156. polarimeters, 104-106. polariscope tube, 157.
sodium lamp, 105, 148. Langen's sweet-water spindle, 48. Laurent's half-shadow polarimeter, 91-93, 101, 102.
saccharimeter, 133-135.
half-wave plate, 91-93.

principle of, 92, 93.
Leach's apparatus for high-temperature polarization, 291, 292. electrolytic apparatus, 407-409.
method
Lead, acetates
of determining
commercial glucose, 291-293
of,
207.
acetate solution, neutral, 207. basic nitrate, clarification with, 218, 219. number of Winton, 517, 518.
precipitate, errors
raffinosate, 740.
due
to,
209.
Hortvet's method of measuring, 516, 517.
removal from solutions (deleading), 276, 277.

saccharates, 681. subacetate, action on rotation of fructose, 217. invert sugar, 185.
amount necessary
sucrose, 216, 217. for clarification, 204.
precipitating action
upon
sugars, 215, 216, 444.
preparation of solutions, 207, 208. use of dry salt for clarifying, 212-215.
Least squares, method
of, 175,
401.
Leffmann and Beam's method
for determining lactose in milk, 253, 254. Leuconostoc mesenterioides, 584, 652, 653, 716. Levan, 615. influence on polarization of sugar products, 654.
Levosin, 615. LevuLan, 615.

/S-Levulin (see Lecalose). Levulinic acid, reaction for hexose groups, 372-374.
yield
from fructose, galactose and glucose, 373.
Levulose
(see
d-Fructose).
INDEX
Lichenin, 578.
Light, dispersion of, 51. effect of kind of on rotation of sugars, 173, 174.
xlvii
polarization
of,
76-82.
for,
sodium, lamps
147-149.
purification of, 149-151.
white, lamps for, 151-153.

Light-filter,
bichromate, 115-117.
Lippich's, 150, 151. of, 76, 77.
Light-wave, theory
Linimarin, 573. Lintner's method for determining diastatic power of diastases, 513. malt, 511-513. preparing soluble starch, 577.
pressure bottle, 439, 440. scale of diastatic power, 512.
Lippich's half-shadow polarimeter, 94-98, 104-106. light filter, 150, 151.
polarizer, 94-98.
principle of, 95-98.
Lobry de Bruyn's method of drying sugars, 25, 26. Lobry de Bruyn and van Ekenstein's method of destroying optical activity
sugars, 303.
of
Lobster
shells,
occurrence of chitin
in,
752.
preparation of d-glucosamine from, 752, 753

Locaose, 631. Lohnstein's fermentation saccharometer, 463, 464. Low's iodide method for determining copper, 411-413.
Lupeose, 748, 749. Lycerose, 630. d-Lyxonic acid, 557.

d-Lyxose, 557.
Main's refractometer table, 64; Appendix, 17. Malt, determination of diastatic power of, 511-513.
preparation of diastase from, 685. process of manufacturing, 684. Malt extracts, action on starch, 685-691.
analysis of, 510, 511. determination of diastatic
power
of,
511-513.
440, 441. restriction of, 690, 691.
preparation
of,
Malt sugar
(see
Maltose).
Maltase, 701, 702. action upon d-glucose, 705. a- and /8-glucosides, 591.
maltose, 705.
Maltobionic acid, 700, 701. Maltobiose (see Maltose).

,
xlviii
Maltodextrin, 577, 686, 706. Maltoglucase (see Maltase). Maltose, 682-705.
INDEX

resorcin, 381.
action of acids on, 701. alkalies on, 701.
enzymes
compounds, 703.
on, 701-702, 705.
maltosates, 703. octacetate, 703.
osazone, 703. configuration, 704. copper-reducing power, 422.
dehydration, 25. determination by copper reduction, 423-426 (see Copper-reduction methods) polariscopic methods, 194-201.
in presence of glucose
and
dextrin, 486-492.
starch conversion products, 486-492, 507, 508. fermentations, 701-703. action of different yeasts, 714.
formation from starch by acids, 697-699. enzymes, 683-696.

hydrolysis of, 701. influence on osazone formation of fructose
and
glucose, 351, 352.
normal weight, 197, 198.

.
occurrence, 682, 683. oxidation products, 700, 701. preparation, 699.

properties, 699, 700. reactions, 700, 701. reducing ratio to glucose, 391, 422. specific rotation, 173-192, 700.
synthesis, 704, 705.

tests, 703, 704.
value of Ventzke degree, 200, 201. variability in reducing power, 402.
Maltose carboxylic
acid, 703.
Manna, 597, 740, 744. Mannans, 593-595.

Mannatetrasaccharide (see Stachyose). Mannatrionic acid, 745.
Mannatrisaccharide, 744, 745. formation from stachyose, 748.
hydrolysis, 744. occurrence, 744.
preparation and properties, 744.
d-Mannite,
INDEX
d-Mannite, formation during fermentation, 653, 654, 764, 765.
occurrence, 597. oxidation by chemical means, 770, 771. bacteria, 771.
xlix
to d-fructose, 617. d-mannose, 596.

properties, 597, 767.
reaction with acetaldehyde, 766.
benzaldehyde, 766-769. borax and boric acid, 765, 766.
formaldehyde, 766.
tribenzal, 770.
1-Mannite, 767.
Mannogalactans, 599. d-Mannoheptite, identity with perseite, 634-635.

properties, 635.
1-Mannoheptite, 635, 768.
d-Mannoheptonic
acid, 634.
rotation of lactone, 774.
d-Mannoheptose, 634, 635.

conversion to d-mannooctose, 639. reduction to d-mannoheptite, 634.
synthesis from d-mannose, 634.
1-Mannoheptose, 635. d, 1-Mannoheptose, 635, 636.
d-Mannonic
acid, 597.
conversion to d-gluconic acid, 775. rotation of lactone, 597, 774. 1-Mannonic acid, 597, 598. conversion to 1-gluconic acid, 775.
d,
1-Mannonic
acid, 598.
resolution
by means
of strychnine salts, 598, 786.
d-Mannonononic acid, 641. d-Mannononose, 641.

d-Mannooctite, 639, 768.
d-Mannooctonic
acid, 639.
d-Mannooctose, 639.
conversion to d-mannononose, 641.
synthesis from d-mannoheptose, 639. d-Mannosaccharic acid, 597.
double lactone of, 597, 780. 1-Mannosaccharic acid, 598. double lactone of, 598.
d-Mannose, 593-597.
resorcin, 381.
action of alkalies upon, 339, 340. conversion to d-mannoheptose, 634.
determination as phenylhydrazone, 469, 470.
INDEX
action of different yeasts, 714.
d-Mannose, fermentation, 596, 597.

formation from methyl mannorhamnoside, 645.
hydrazone, 597.
mutarotation, 596. occurrence, 593-595.
oxidation, 597. osazone, 353, 354, 597.
preparation, from Carob beans, 596. ivory nuts, 595.

properties, 596, 597. reduction, 596, 597.
specific rotation, 596.
synthesis, 596. tests, 597.
1-Mannose, 597, 598.

d,
action of yeasts upon, 714. 1-Mannose, 598. Maple sugar, composition of ash, 519. lead
number
of,
518.
volume
of lead precipitate from, 517.
Maquenne's block, 357-359.

test for methylfurfural, 377, 378.
Maquenne and Roux's theory

Marc, determination, 228, 229.
of diastatic conversion, 687-689.
variation of content in beets, 246. Marcker's diastase method for determining starch, 440, 441. Mashing at high and low temperature, 690.
Massecuite, 646. determination of moisture
in,
18-20.
methods
for polarizing, 205, 206.

i-inosite from, 760.
Mazun,
Meissl's
714.
Meat, preparation of
method for determining invert sugar, 423; Appendix, 38. Meissl and Killer's method for determining invert sugar in presence
431.
of sucrose, 430,
Meissl and Wein's method for determining invert sugar in presence of sucrose, 428430.
Melassigenic action of
salts,
648-650.
Geerligs's theory of, 650. in beet molasses, 649.
cane molasses, 650.

Melezitose, 740-742.
fermentation, 742. hendecacetate, 742.

hydrolysis, 741.
occurrence, 740, 741. preparation, 741.

properties, 741.
reactions, 741.'
INDEX
Melibiase, 723.
li
Melibiose, 721-724.
absorption
of,
by bone black,
284, 285.
fermentation, 723. formation from raffinose, 737, 738,

hydrolysis,
by
acids, 723.
melibiase, 723. occurrence, 721.
preparation from raffinose, 721, 722.

properties, 722.
reactions, 722, 723. synthesis, 724.
tests, 723, 724. Melibiotite, 722. Melitriose (see Raffinose). Melting points of hydrazones
and osazones, 356-359; Appendix, 90. comparison of methods for determining, 359.
determination by capillary tube, 356, 357. Maquenne's block, 357-359.
variability in, 359, 360.
Meniscus, adjustment
of,
167.
1-Menthylhydrazine, 362. use of in resolving d, 1-sugars, 362, 551. Mercaptals, 367, 368. Mercaptans, reactions of sugars with, 367, 368. Mercury, acid nitrate solution for clarifying, 252, 447.
iodide solution for clarifying, 252. Knapp's alkaline cyanide solution for determining reducing sugars, 338, 435, 436.
Sachsse's alkaline iodide solution for determining reducing sugars, 436, 474.
Mesoerythrite
(see i-Erythrite).
Metal polariscope tubes, 157-159.

with jacket, 158, 159.
Metallic salt solutions for testing sugars, 334-339 (see under Bismuth,
Copper.
Mercury and
miscellaneous solutions, 339.
Silver),
Metabolism
of sugars in plants, 534.
Metapectic acid, 601. Metapectin, 601. Metaraban, 547. Metarabin, 548. Metasaccharins, 587, 604.
Methyl alcohol, use of, in separating raffinose and Methyl a- and /3-glucosides, 590, 591. mannorhamnoside, 645.
Methylarbutin, 571.
Methyldioses, 535, 536.
Methylerythrite, 543, 767. Methylfurfural, absorption spectra, 384-386.
color reactions, 377, 378.
sucrose, 734.
Hi
INDEX
Methylfurfural, determination, 456-459. in presence of furfural, 458, 459.
formation from methylpentoses and methylpentosans, 377, 378. method for determining methylpentoses and methylpentosans, 456459; Appendix, 89. phloroglucide, 457. factors for converting to methylpentoses pentosans, 457.
precipitation with barbituric acid, 457. phloroglucin, 456, 457.
and methyl-
reaction for methylpentose groups, 377. yield from methylpentoses, 377.
Methylglycerose, 539.
Methylglycolose, 535.
Methylglyoxal, 378, 537, 539.
Methylheptoses, 637, 638. Methylhexoses, 631-633.

Methyloctoses, 640. Methylpentosans, determination
pentoses).
of,
456-459; Appendix, 89
(see also
under Methyl-
Methylpentose-hexose saccharides, 644, 645, 731, 732. Methylpentoses, 563-570. color reactions, 384-386.
determination, 456-459; Appendix, 89. by method of Tollens and Ellett, 456-458.
Haywood's modification, 458. in presence of pentoses, 458, 459. methylfurfural reactions, 377, 378 (see also under Methylfurfural). Methylphenylhydrazine, 346.
reaction for d-fructose, 621, 622. ketoses, 354.
use of in determining d-fructose, 470.

Methyltetroses, 543.
Methyltrioses, 539. Metric cubic centimeter, 28.
normal weight, 113, 114, 163, 203.

standard in saccharimetry, 113-115. Metric solution scale, 163.
Microorganisms, action upon samples, 13, 14. Midzu ame, analysis of, 491. Milk, clarification of, 447. determination of lactose in, 252-254.
percentages of lactose in different kinds polarization of, 252-254.
of,
708.
Milk sugar
(see
Lactose).
of cane juice, 232.
Milling, effect
on composition
Mitscherlich's polariscope, 85, 86.
Mixtures of sugars, analysis
of,
279-286, 472-493.
by combined
polariscopic methods, 472, 473. reduction methods, 473, 474.
INDEX
liii
Mixtures of sugars, analysis of, by combined polariscopic and reduction methods, 475-493. determinations of two sugars in, 475-483.
arabinose and fructose, 482.
fructose
xylose, 482, 483. galactose, 481.
and
glucose, 477-479.
and glucose, 480, 481. glucose and sucrose, 279.

galactose
lactose
and
xylose, 300, 301. sucrose, 280, 281.
methylpentoses and pentoses, 458, 459. raffinose and sucrose, 282-285. determinations of three sugars in, 484-492.
dextrin, glucose
fructose, galactose
and maltose, 486, 490. and sucrose, 484, 485. glucose and sucrose, 485, 489.
in,
determinations of four sugars formulae for analyzing, 477. racemic, 532.
492, 493.
resolution of, 361, 362, 784-787.
Mohr
cubic centimeter, 28.
normal weight, 113, 163.

standard in saccharimetry, 113.
Mohr's
specific gravity balance, 40-42.
Moisture, absorption of by raw sugars, 7-9. determination by drying in air, 16-20.
vacuum, 20-24. on sand, 19. pumice stone, 18>
19.
method
of A. O. A. C., 16-18. Browne, 23, 24.
Carr and Sanborn, 21-23. International Commission, Lobry de Bruyn, 25, 26.
Pellet, 19, 20.
16.
in
commerical dextrin, 508. fructose and its products, 20-24.

glucose, 25.
lactose, 25. maltose, 25.
raw
sugars, 15-18.
sirups, molasses, etc., 18-26. starch products, 26.
estimation from refractive index, 50-75. specific gravity, 27-49.
evaporation of from raw sugars, 7-9. Molasses, calculation of composition and purity in raw sugars, 506. comparison of methods for determining solids in, 69. determination of moisture in, 18-25.
liv
INDEX
by clarification, 69, 70. dilution with water, 66-68. sirup, 68, 69.
specific gravity, 38.
Molasses, determination of refractive index, 66-70.
dextrorotation at 87 C. after inversion, 296.

effect of clarifying agents
on polarization, 224, 225.
methods
for polarizing, 205, 206. occurrence of glutose in cane-, 629.
preparation of raffinose from beet-, 734, 735. Molecular depression of freezing point, 329.
elevation of boiling point, 331, 332.
heat of combustion, 318.
rearrangements of sugars, 303, 625, 626, 628, 629. sugar acids, 775. weight determinations of sugars, 322-332.
by
boiling point method, 331, 332.
freezing point method, 327-331.
osmotic pressure, 322-324. plasmolysis, 324, 325.
Molybdates, color reactions of sugars with, 682. influence of, on rotation of sugar alcohols, 766. Monnier's formula for calculating rendement, 498.
Monosaccharides, 535-642.
classification, 528.
and acids, 529, 530. variability in reducing power, 400.

relationship to alcohols
Morfose, 630.
Moulds, non-inverting, 651. occurrence of chitin
in,
752.
Mounting
Mucic
polariscopes, 169, 170. polariscope tubes, 154.
acid, 605.
configuration, 605.
conversion to allomucic acid, 781.
dehydromucic
lactose, 712.
acid, 605, 781.
formation from galactose, 364, 459, 460, 604, 605.

properties, 605. reaction for galactose group, 364, 459, 460, 604, 605. reduction, 783.
yield
from galactose, 459.
Mucins, 751, 752.
Mucor
circinelloides, 651.
Mucose, 631.
Multirotation
(see
Mutarotation)
Munson and Walker's method
for determining d-glucose, invert sugar, lactose maltose, 426; Appendix, 66.
and
method
for determining invert sugar in presence of sucrose
432; Appendix, 66.
INDEX
Munson and Walker's method
Muscovado
for preparing asbestos, 406.
1?
sugar, composition of ash, 519. lead number of, 518.

of chitin in, 752.
Mushrooms, occurrence
trehalose in, 718.
Mushroom
sugar (see Trehalose). Mutarotation, 187-193.

influence of acids on, 189, 190. alkalies on, 190.
salts on, 190. solvent on, 190, 191.
temperature on, 188. occurrence during inversion, 671-673.

of d-glucose, 189.
lactose, 709-711.
theories of, 191-193. velocity of, 188.
Mycose
(see
Trehalose).
Myrosin, 573.
Myrticolorin, 599.
a-Naphthol, absorption spectra of sugars with, 379.

color reaction of sugars with, 341, 378, 379. test for sucrose, 341, 681.
Naphthoresorcin, absorption spectra of glucuronic acid with, 383-385. pentoses with, 383-385. sugars with, 381. test for glucuronic acid in urine, 383-385.
Nasini and Villavecchia's concentration formula for specific rotation of sucrose, 176.
Naphthylhydrazine, 347. Nectar, composition of, 616.
Net
analysis, 498.
Neutral polarization of inverted solutions, 271. New York Sugar Trade Laboratory, constant temperature cabinet, 169. methods of polarization, 202-205, 261.
refrigerating equipment, 261, 262.
New York
Sugar Trade method of sampling sugar,
6.
Nicol prism, 81-84. Nitric acid, inverting power of, 663. oxidation of sugar alcohols with, 770, 771.
sugars with, 364.
p-Nitrophenylhydrazine, 347. Nomenclature of sugar acids, dibasic, 778. monobasic, 773.
sugar alcohols, 766.

sugars, 528, 531, 532.
Non-inverting yeasts and moulds, 651.
Nonoses, 640, 641. Non-reducing sugars, reactions

Non-sugar, 496.
of,
386, 387,
Ivi
INDEX
Non-sugar, organic, 496.
"Nori,"565, 606.
Normal juice, 497. Normal weight of sucrose
for saccharimeters, 112-119.
for
French scale, 112, 113. Ventzke or German Scale, 113-115.

metric
cc.
cc.
Mohr
U.
S.
standard, 113, 114, 163. standard, 113, 163.
Coast Survey standard, 114, 115.
Normal weights
of sugars, 197-201.
definition, 197, 199.
methods
of calculating, 197, 198. tables of, 197, 198. use of a single weight for all sugars, 200, 201.
Norrenberg's polariscope, 78-80. Noyes's modified diastase method for determining starch, 442. Nucite (see i-Inosite).
Nucleic acids, occurrence of pentose group Nutritive salt solution for yeast, 299.
in,
558.
Nylander's bismuth solution, 338.
Oak sugar (see Quercite). Octoses, 638-640.

Optical activity of sugars, methods of destroying, 302-306. relation to asymmetric carbon atom, 530.
Optical inactivity of sugars, 531. Optically inactive sugar from sucrose, 659. Organic matter, determination of, 496.
Organic non-sugars, determination of, 496. Osazones of sugars, analysis of, 370, 371.
conversion into osones, 354, 355.
elementary composition, 371.

îdentification, 356-361, 370, 371. limitation of reactions for, 353, 354.
melting point
(see
Melting points).
purification, 353.
rotation of, 360, 361. table of formulae, descriptions, melting points, Appendix, 90.
yield and time of formation, 350-353. process, 649, 650. Osmotic pressure of sugar solutions, 321-332.
and
solubilities,
Osmose
application to molecular weight determinations, 324.
determination by Pfeffer's method, 322-324.

plasmolysis, 324, 325. relation to boiling point, 326, 327. freezing point, 326, 327.
gas pressure, 323.
vapor pressure, 326, 327.

Osones, formation from osazones, 354, 355.
INDEX
Ost's copper bicarbonate method for determining reducing sugars, 433, 434. O'Sullivan's copper reducing factors, 421, 422.
Ivii
solution factors, 31.
O'Sullivan and Tompson's yeast method of inversion, 274.

Ogilvie's modification, 274.
Oven, Carr's vacuum, 22, 23.

Soxhlet's, 16, 17.
Wiesnegg's, 17, 18.

Oxalic acid, inverting power of, 663. use in Clerget method, 273, 274 Oxalic fermentation, 585. Oxidizing agents, action upon sugars, 363, 364. Oxime reaction of sugars, 364, 365.
Oxycellulose, production of furfural from, 376.
Oxymethylfurfural, formation from d-fructose, 620.
Oxymethyltetroses, 544.
Pancreatin, 693-696 (see under Conversion of Starch). Paper-stock, determination of pentosans in, 456.
Paradextran, 578.
Parapectic acid, 601.
Parasaccharose, 729. Pasteur's methods of resolving racemic mixtures, 785-787. researches upon alcoholic fermentation, 582.
tartaric acid, 784-787. Pavy's volumetric method for determining reducing sugars, 395-397. Payen's method for determining crystal content, 499.
Pectase, 601. Pectic acids, 601. Pectinase, 601.

Pectins, 600-602. Pectose, 601.
Pectosinase, 601.
Pellet's
drying capsules, 19. method of aqueous digestion, 239, 240. with cold water, 239. hot water, 240.
determining moisture, 19, 20. tube for continuous polarization, 158, 159. Pellet and Lemeland's method for destroying optical activity of reducing sugars,
305, 306.
Pellin's polarimeter, 102, 103.
saccharimeter, 135. Penicillium glaucum, selective action
on
d, 1-acids, 787.
Pentosans, methods of determining, 449-456; Appendix, 83 (see also under Pentoses). occurrence, properties, etc. (see under Araban and Xylan),
percentages in paper and paper stock, 456. Pentose-hexose saccharides, 643, 644. Pentoses, 545-562.
apparatus for determining, 450, 451.
Iviii
INDEX
Pentoses, color and spectral reactions, 381-386. with naphthoresorcin, 383-385. orcin, 382, 383.
phloroglucin, 381, 382, 384.
determination from yield of furfural, 449-456.
by barbituric acid method,
454.
phloroglucin method, 451, 452. titration with bisulphite, 454, 455.

factors for calculating
from phloroglucide, 452.
furfural reaction for, 374-377 (see under Furfural). Krober's table for determining, 451; Appendix, 83.
limitations of methods for estimating, 375-377, 452, 453. Permanganate volumetric method for determining copper, 410, 411. Perseite, 634, 635 (see also d-Mannoheptite) conversion to perseulose, 637
.
dibenzal, 770.
occurrence, 634, 635. properties, 635, 767.

Perseulose, 637. Peters' s saccharimeter, 138.
Peter's electrolytic method for determining copper, 409, 410. iodide method for determining copper, 413, 414.
Pfeffer's researches
Pfliiger's
on osmotic pressure
of sucrose solutions, 322-324.
method
of determining glucose, 419;
Appendix, 33.
glycogen, 443.
Pharbitose, 729. Phaseolunatin, 573.
Phaseomannite
(see i-Inosite).
Phenols, color reactions with sugars, 341, 378-386. reactions of sugars with, 368.
Phenylhydrazides
(see Hydrazides). Phenylhydrazine, reaction with acids, 777, 782. reaction with sugars, 345-362
(see also
under Hydrazones).
substituted derivatives
346, 347, 361, 362. use in determining d-mannose, 469, 470.

of,
Phenylhydrazones
Phlein, 615.
(see
Hydrazones)
Phloridzin, 571, 578.
Phloroglucide, furfural, 375, 451, 452. methylfurfural, 456, 457.

Phloroglucin, color reactions with pentoses, 381, 382, 384. purification of, 451.
use in precipitating furfural, 451, 452.

methylfurfural, 456, 457.
Photosynthesis, 533.
Phytase, 763.
Phytin, 762, 763.
Pinite, 758, 759.
occurrence, 758, 759. preparation of d-inosite from, 758.
INDEX
Finite, properties, 759.
lix
Pipette, Kruger's automatic, 240, 241. Sachs-Le Docte automatic, 243.
Spencer's sucrose, 205.

viscosity, 307.
Plant tissues, distribution of water " Plaque type," 135.

Plasmolysis, 324, 325.
in,
230-232.
application to molecular weight determinations, 325.

Plate, concentric half-wave, 93. Laurent's half-wave, 91-93.
Savart's 98.
double quartz, 86, 88. Plates, standard quartz, for calibrating polariscope scales, 119, 120, 135. Polarimeters (angular degree Polariscopes), 76-107 (see also under Polariscopes).
Soleil's
apparatus of Biot, 84, 85.

Jellet, 89.
Landolt, 104-106.
Laurent, 91-93, 101, 102.

Lippich, 94-98, 104.
Mitscherlich, 85, 86. Norrenberg, 78-80.
Pellin, 102, 103.
Robiquet, 86-88. Wild, 98-101.

construction
of,
82-101.
factor for converting readings into sugar degrees, 145. half -shadow instruments, 89-98.
description of
scales
modern
types, 101-106.
and method
of reading, 85-87.
tint instruments, 86-88.
verification of scale readings, 106, 107.
Polariscopes, 76-145 (see more especially under Polarimeters accessories of, 146-171.
and Saccharimeters)
analyzer
of,
82-84.
angular degree
(see Polarimeters). cabinets for (see Cabinet), care of, 169-171.
field of vision (see Field),
illumination
of,
146-153.
mounting
of,
169, 170.
polarizer of, 82-84.
quartz-wedge (see Saccharimeters). sugar degree (see Saccharimeters). theory of, 76-101. tubes for (see Tubes). Polariscopic methods, employing direct polarization, 194-262. double polarization, 263-286. special processes, 286-306.
for analyzing sugar mixtures, 472, 473, 475-493.
lx
INDEX
Polariscope methods for determining velocity of inversion, 661, 662. (see under Filter-press cake, Honey, Milk, Molasses,
Raw
sugars, Sugar beets, etc., for particular methods).
Polaristrobometer of Wild, 98-101. Polarization at constant temperature, 261, 262.
equipment
Polarization at high temperature, 287-299.
for, 169,
262.
for determining
commercial glucose, 289-296.

fructose, 296-299.
invert sugar, 287-289.
method
of
Chandler and Ricketts, 289-291.

Leach, 291-293. Wiley, 296-298.
Polarization of light, 76-84.
by double
theory
Polarizer, 82-84.
of Jellet, 89.
of,
refraction, 80-84.
reflection, 78-80.
76-84.
Cornu's modification, 89-91.

of Lippich, 94-98. of Schmidt and Haensch, 89.
Polysaccharides, 528, 574-579. Populin, 571.

Precipitation of sugars by basic lead salts, 215, 216. Precipitate error, in polarizing milk products, 253, 254. sugar products, 209, 215.
methods
of correcting, 209-215.
by Home's method,
212, 213. Sachs's method, 210, 211. Scheibler's method, 209, 210.
Press, hydraulic, for laboratory use, 227, 228. " Sans Pareille," 239.
Pressure bottle, Lintner's, 439, 440. Pressure methods for dissolving starch, 439, 506. Pressure, osmotic (see Osmotic pressure).
Pfibram's sodium lamp, 148. Prism, Glan, 82.

Jellet-Cornu, 89. Nicol, 81, 82.
Protagon, 602.
Prulaurasin, 572, 573. Prunose, 560.
Pseudinulin, 615. Pseudofructose, 629.
Pseudostrophanthobiose, 729.
Ptyalin, 693. Purification of
bone black, 219.

osazones, 353.
sirups in preparing sugars, 550.
INDEX
Purification of sodium light, 149-151. Purity, coefficient of, 494, 495.
bd
determination in molasses of raw sugars, 506. Purity of reagents, influence on copper reduction, 417.
Pycnometer, Boot's, 38.

types
of,
36-39.
Pyromucic
acid, 782.
Quadruple
field, 97, 98.
Qualitative methods for examining sugars, 333-387. Quartz, rotation of, compared with sucrose, 116, 117. temperature coefficient for rotation of, 126, 127.
Quartz plates, for verifying polariscope "plaque type," 135. Quartz wedge, compensation, 108-112.
scales, 119, 120.
double, 110-112. single, 108-110.
Quebrachite, 759.
Quercinite, 762. Quercite, 756, 757.
occurrence, 756.
properties Quercitrin, 563.
and
tests, 757.
Quinovin, 568.
Quinovose, 568, 569.
Racefoliobiose, 730. Racemic mixtures, 532.
resolution of d, 1-acids, 784-787.
by
crystalline form, 785, 786. optically active bases, 786, 787. selective fermentation, 787.
resolution of d, 1-sugars, 551, 593, 598, 607, 623. by optically active hydrazines, 361, 362. selective fermentation, 787.
Radiation correction in calorimetry, 316. Raffinose, 732-740.

resorcin, 381.
action of emulsin on, 737, 738. invertase on, 737, 738.

calorific values, 318, 319.
compounds, 738-740.
hendecacetate, 738, 739.
octobenzoate, 739.
raffinosates of barium, calcium, strontium, lead
and sodium,
739-740.
configuration, 738, 740.
dehydration, 25.
Ixii
Raffinose, determination,
INDEX
by double
polarization, 281-286.
Creydt's method, 282. Herzf eld's method, 282, 283.

error
reliability of
from bone-black absorption, 284, 285. methods, 285, 286. temperature correction for, 283, 284.
fermentation, 738. formation of melibiose from, 721, 722. hydrolysis, by acids, 736, 737.
enzymes, 737, 738.

influence
on
crystalline
form
of sucrose, 735, 736.
formation of d-glucose-osazone, 352. molecular weight determination by plasmolysis, 325.
normal weight
of,
197, 198.
occurrence, 732, 733. preparation from beet molasses, 734, 735. cotton-seed meal, 733, 734.
properties, 735.
reactions, 736-738.
separation from sucrose, 734, 735.

solubility, 735.
specific rotation, 173, 174, 736.
tests, 740.
value of Ventzke degree, 200, 201. Raoult's method for determining molecular depression of freezing point, 327-331. Rapp-Degener method of alcoholic digestion, 238.
Rate
of inversion, 660-662.
Raw
sugars, clarification of solutions, .204, 207-215, 276-278. composition, 259, 260.
deterioration of samples, 14.

in, 15-18, 65. agents on polarization, 224. polarization, 201-205.
determination of moisture
effect of clarifying
method
sampling, method of
of
New York
rules of International
New
U.
S.
Sugar Trade, 202-205. Commission, 201, 202. York Sugar Trade, 5, 6. Treasury Dept., 5, 6.
shaker for dissolving, 203, 204.

Reciprocals, table "Redo," 221.
of,
398; Appendix, 101.
Reducing action
law of, 400, 401. sucrose on Fehling's solution, 427, 428. agents, reaction of sugars with, 362, 363.
of sugars,
power, relative copper, 421-423.

variability in, of disaccharides, 402. monosaccharides, 400.
ratio of sugars, glucose equivalent, 391, 421.
Reducing sugars, determination
maltose equivalent, 422. (see Copper reduction methods),

of,
glucose equivalents
427, 476.
INDEX
Reducing sugars, precipitation by basic lead
salts, 215, 216, 444.
Ixiii
reactions, 333-386. volume of Fehling's solution reduced by, 391. Reduction methods, combined, for analyzing sugar mixtures, 473-475.
Reduction tables of sugars, calculation

Refining of
of,
401.
raw
sugar, 646, 647.

(see
Refining value, 498
Rendement).
Reflection, principle of total, 52, 53. Refraction, law of, 50, 51.
Refractive index of sugar solutions, 50-75.

calculation of dissolved solids from,
clarification of solutions for, 69, 70.
51-75.
determination by Abbe's refractemeter, 53-70.
immersion refractometer, 70-75.

dilution of solutions for, 66-68. influence of impurities on, 66-70.
temperature on, 58, 59. relation of to specific gravity, 62. Tischtschenko's method of determining, 68, 69.
Refractometer,
Abbe
(see
Abbe
(see
refractometer)
immersion
Immersion refractometer).
table of Geerlig's, 65; Appendix, 22.
Refractometer tables for sugar solutions, 61-65.
Hubener, 74; Appendix, 24. Main, 64; Appendix, 17.

Schonrock, 64.
Stanek, 64; Appendix, 21.
Stolle, 62.
Strohmer, 61.
Tolman and Smith,
62, 63.
Refrigerating machine for constant temperature polarization, 262. Reischaur and Kruis's method for determining glucose, 398, 399; Appendix, 27. Rendement, methods for calculating, 498.
Reputed cubic centimeter,
28.
Resorcin, absorption spectra of sugars with, 381, 384. color reactions of sugars with, 380, 381.
test for ketoses, 380.
Restriction of malt extracts, 690, 691.
Reversion products, formation from starch, 488, 697. Revertose, 704, 730.
Rhamninase, 731. Rhamninite, 731. Rhamninose, 731, 732.

hydrolysis, 732.
occurrence and preparation, 731.

oxidation, 731, 732.
properties
and
reactions, 731, 732.
Rhamninotrionic acid, 732.

Ilhamnite, 565, 767.
Rhamnodulcite
(see
Rhamnose).
Ixiv
Rhamnoheptonic
acid, 637.
INDEX
Rhamnoheptose, 637, 638.

conversion to rhamnooctose, 640.
a-Rhamnohexite, 631, 768.
a-Rhamnohexonic
acid, 631.
/S-Rhamnohexonic acid, 632.

a-Rhamnohexose, 631, 632.

conversion to rhamnoheptose, 637. /3-Rhamnohexose, 632.
Rhamnonic
acid*,
565.
Rhamnooctonic
acid, 640.
Rhamnooctose, 640. Rhamnose, 563-565.

resorcin, 381.
calorific values, 319.
conversion to isorhamnose, 568, 777.
rhamnohexose, 631.
determination, 456, 457; Appendix, 89. fermentation, 565.
action of different yeasts, 714
formation from glucosides, 563, 564.
mannorhamnoside, 645.
rhamninose, 732.
563, 564. modifications, 564.
glucosides
of,

occurrence, 563, 564. oxidation with bromine, 363, 565. preparation from quercitrin, 564.
influence of alcohol on, 182, 565.

tests,
565
(see also
under Methylpentoses)
yield of methylfurfural from, 377. Rhamnosan, 457.
determination, 456, 457; Appendix, 89
(see also
under Methylpentosans)
a-Rhodeohexose, 632.
0-Rhodeohexose, 633.
Rhodeonic
acid, 567, 568.
Rhodeose, 566-568. conversion to rhodeohexose, 632, 633.

occurrence, 567.
preparation from convolvulin, 567.
INDEX
Rhodeose, properties, 567. racemic combination with fucose, 568.
tests, 567, 568. 1-Ribonic acid, 558, 559. conversion to 1-arabonic acid, 775. rotation of lactone, 774.
Ixv
d-Ribose, 558.
1-Ribose, 558, 559.
d, 1-Ribose,
formation from adonite, 559.
Robiquet's polariscope, 86-88.

Rolfe's
for calculating composition of acid hydrolyzed starch products, 507. researches on acid conversion of starch, 698, 699. Rolfe and Faxon's method of drying starch products, 26.
method
"Rongalite," 222.
Ross's method of testing for unreduced copper, 393, 394. Rotation dispersion of sugars, 115, 173, 196.
Rotation, specific
(see Specific rotation).
Ruberythric acid, 572.

Saccharan, 467, 656.
Ehrlich's colorimetric
method
for estimating, 467.
Saccharate, polarization Saccharates, 676-681.
of,
in filter-press cake (see under Filter-press cake).
barium monosaccharate, 680. calcium bisaccharate, 677.

monosaccharate, 677.
trisaccharate, 678.
lead saccharate, 681.
potassium saccharate, 676.
sodium saccharate, 676.

strontium bisaccharate, 679, 680. monosaccharate, 678, 679. Saccharic acid, 587-589.
acid lactone
of,
779, 780.
test for d-glucose groups, 587-589.
Saccharides
under Mono-, Di-, Tri-, Tetra-, and Polysaccharides) hydrolytic methods of determining higher, 436-443. Saccharimeters (Quartz-wedge polariscopes), 108-145.
(see
apparatus of Bates, 139-143. Chandler and Ricketts, 290, 291.

Ducboscq-Pellin, 135.
Fric, 138, 139.
Jellet-Cornu, 133.
Laurent, 133-135.
Peters, 138.
Schmidt and Haensch, 136-138.

Soleil-Duboscq, 132.
Soleil-Ventzke-Scheibler, 131.
Stammer,
construction
of,
144.
108-112.
INDEX
Saccharimeters, conversion factors for scales of, 145. of readings to angular rotations, 145, 196. weights of sugars, 199-201.
correction of readings for concentration, 118, 119.
temperature, 255-262. graduation of scales, 117-119. for variable temperatures, 129, 130. half-shadow instruments, 132-145.
normal weight (see Normal weight), quartz-wedge compensation (see Quartz-wedge),

scales, 111, 112.
method
of reading, 111.
temperature corrections, by method of Browne, 258-261. U.S. Treasury Dept., 256, 257.
Wiley, 256.
.
error of methods, 257-261. for beet products, 258, 260.
cane products, 258, 259, 261. temperature, effect on scale readings, 126-130.
tint instruments, 131, 132.
use of bichromate light
filter,
115-117.
verification of scale readings, 119-126.
by
control tube, 122-125.
"hundred polarization,"
quartz plates, 119, 120.
sucrose, 121, 122.
125, 126.
with magnified
Saccharimetry, special methods
technical
scale, 143, 144.
variable sensibility, 139-143.

of,
287-306.
of,
methods
201-286.
Saccharin, 586, 587. Saccharinic acid, 586, 587.
Saccharometer, Einhorn's fermentation, 462. Lohnstein's fermentation, 463, 464.
Saccharomyces apiculatus, 651.

cerevisise, 582, 714.
ellipsoideus, 714.
Marxianus, 704, 705, 714. membransefaciens, 714.

octosporus, 651. productivus, 714.
Saccharose
Sachs's
(see Sucrose).
of determining lead precipitate error, 210, 211. Sachs-Le Docte automatic pipette, -243. process of water digestion, 242-244.
method
Sachsse's
method
for determining
reducing sugars with mercuric iodide solution,
Salep mannan,
Salicin, 571.
436, 474. for determining starch, 439. 594.
INDEX
Saline quotient, 496. Saliva, determination of diastatic
Salts, influence
Ixvii
power
of,
515
on activity
of diastase, 691.
pancreatin, 694. inverting power of acids, 665. rotation of reducing sugars, 184, 185.
inverting action Sambunigrin, 572.
upon
sucrose, 183, 184. sucrose, 666-668.
Sampler, Coomb's drip, 10, 11. Samples, changes in composition
of,
12-14.
of moisture, 12, 13.
by absorption and evaporation

action of enzymes, 13.
microorganisms, 13, 14.

deterioration
of, 13, 14.
mixing
of, 5, 9.
of, 14.
preservation
segregation of molasses
in, 9.
sugar crystals in, 11, 12. Sampling of juices, molasses and sirups, 10, 11.
raw
sugars, 3-10.
change in moisture content during, 6-9.

introduction of trash during,
9.
method
of
New York
U.
S.
Sugar Trade, 6. Treasury Dept., 5, 6
triers for, 4-6.
sugar and sugar products, 3-14.

errors in, 6-12.
sugar beets, 225-227.
"Sans Pareille"
press, 239.
Saporubrose, 631. Savart plate, 98. Scale, metric solution, 163.

Scales of polarimeters, 85-87.
method of reading, 85-87.

verification, 106.
zero-point determination, 106.
snccharimeters, 110-130. conversion factors for, 145.
German
French sugar scale, 112, 113. or Ventzke scale, 113-115.

for metric c.c., 113, 114. Mohr c.c., 113.
graduation, 117.
magnified, 143, 144,
method
of reading, 111.
verification,
(see also
119-126 under Polarimeters and Saccharimeters).
Scammonose, 631.
Scheibler's elution pj ocess, 67.
Ixviii
Scheibler's
INDEX
method
of alcoholic extraction, 233-235.
determining crystal content, 499-501. double dilution, 209, 210. "hundred polarization," 125, 126
saccharimeter, 131. specific gravity tables for sucrose, 29. strontium process, 679, 680.
Scherer's test for inosite, 758. Schiff's reaction for furfural, 374.
Schmidt and Haensch polariscope tube, 157.

polarizer, 89.
saccharimeters, 136, 137, 153. Schmitz's concentration formula for rotation of sucrose, 118, 176. table for correcting saccharimeter readings, 118.
Schonrock's formula for temperature coefficient of saccharimeters, 120. table of refractive indices of sucrose solutions, 64.
Scillin,
615.
Secalose, 746. Seliwanoff's resorcin test for d-fructose
and
ketoses, 380, 384, 619.
Semicarbazone reaction of sugars, 366. Seminose (see d-Mannose). Shaking machine for dissolving sugars, 203, 204. Sherman's researches on pancreatin, 693-696.
scale of diastatic power, 514, 515.
Sherman, Kendall and Clark's method for determining diastatic power, 513-515. Sherman and Williams's results on time of osazone formation, 351-353.
Sidersky's specific gravity tables, 30. Sieben's method for estimating fructose, 470, 471 Silver solution of Tollens, 337, 338.
Simple sugars
Sinalbin, 573.
(see
Monosaccharides).
108, 109.
Single
wedge system,
Sinigrin, 573.
Sinistrin, 615.
methods for polarizing, 205, 206. purification of, in separating sugars, 550. Skimminose, 631.'
Sirups,
Sodium
light, 77, 116, 173.
lamps
for,
147-151.
purification of, 149-151. wave length of, 150.
Sodium hydrosulphite, as a
raffinosate, 739.
decolorizer, 221.
saccharate, 676. sulphite, as a decolorizer, 278. Solanose, 631.

Soldaini's copper solution, 337, 432.
Soleil's
double-quartz plate, 86-88. quartz wedge compensation, 108.

saccharimeters, 131, 132.
INDEX
Solubility of sucrose in water, at different temperatures, 649. influence of salts on, 648-650. Soluble matter, determination in commercial dextrin, 509.
Ixix
Soluble starch, determination in commercial dextrin, 509. Lintner's method of preparing, 577.
Solution
by weight, flasks for, 164. Solution factors, 31, 32. use in analysis of starch-conversion products, 487. Solution scale, metric, 163.
Solutions, sugar, boiling point of, 331, 332. concentration of, 448.
freezing point
of,
327-331.
321-327.
plant substances, 445, 446.
isotonic, 326, 327.
osmotic pressure
preparation
of,
of,
from animal substances, 447.
refractive index of, 50-75.

specific gravity of, 27-48.
of, 326, 327. viscosity of, 307-313. Solvent, influence on rotation of sugars, 181, 182. Sophorin, 563.
vapor pressure
d-Sorbinose
(see d-Sorbose). d-Sorbite, dibenzal, 770.
formation by reducing d-fructose, 619.

occurrence, 624. oxidation by Bacterium xylinum, 624. properties, 767.
reaction with benzaldehyde, 769.
1-Sorbite, 627, 767.
d-Sorbose, 623-625.

resorcin, 381.
fermentation, 625. occurrence, 623, 624.
preparation from d-sorbite, 624.

specific rotation, 181, 625.
tests, 625.
1-Sorbose, 625-627.
properties, 626.
synthesis from d-galactose, 625, 626.

tests, 627.
d, 1-Sorbose, 627.
Sorbose bacterium
(see
Bacterium xylinum).
Soxhlet's autoclave, 439, 440.
drying oven, 16, 17.

extractor, 234.
^'uller's modification of, 234.
Ixx
Soxhlet's
INDEX
method
for analyzing sugar mixtures, 473, 474.
determining lactose, 424; Appendix, 42. reducing sugars, 389-391.

modifications
Specific gravity balance, 40-42.
bottles, 36-39.
of,
391, 392.
Specific gravity of
impure sugar solutions, 35, 36.

lead precipitates, 211, 212.
starch-conversion products, 31, 487. sucrose solutions, 28-34.

influence of impurities on, 35, 36.
temperature on, 30, 31. relation to refractive index, 62. table of Balling, 29.
Brix, 29; Appendix, 6. Gerlach, 29.
German Imperial Commission,

Appendix,
1.
30;
Scheibler, 29. Sidersky, 30.
sugar solutions, 27-49.

calculation of solids from, 27-36. by solution factors, 31, 32.
tables, 28-31. errors in, 35, 36.
methods
Specific heat of
of determining, 36-49.
combustion
(see Calories).
Specific rotation of lactones, 774, 775.
determination of configuration from, 774, 775.

Specific rotation of starch conversion products, 31, 507.
calculation of ingredients from, 507, 508.

Specific rotation of sugars, 172-193.
calculation of, 172, 173, 194. determination of concentrations from, 194.
normal weights from, 197, 198.

effect of acids on, 185, 186.
concentration on, 174-177. foreign optically active substances on, 186, 187. kind of light on, 173.
mineral impurities on, 183-185.

solvent on, 181, 182. temperature on, 178-181.
influence of mutarotation on, 187-193. Specifications for sugar flasks, 166-168. Spectra, absorption, for identifying sugars, 342-345, 378-386. diagram of characteristic, 384.
methods
(see
of studying, 344, 345, 378-386. under Glucuronic acid, Methylpentoses, Pentoses, and the different sugars
for individual spectral reactions).
INDEX
Ixxi
Spectroscope, direct-vision, 342-344. 378-386. applications to study of color and spectral reactions, 344, 345, 206. sucrose pipette, 205, Spencer's
Stachyose, 747, 748.

fermentation, 748. hydrolysis of, 748.
occurrence, 747. preparation, 747. properties, 747, 748.
Stammer's colorimeter, 467-469.

saccharimeter with magnified scale, 144. Standardization of hydrometers, 43-45.
polariscope tubes, 155, 156. refractometers, 59-61, 72-74. saccharimeter scales, 119-126.
sugar flasks, 166-168. Stanek's zinc nitrate method for decomposing saccharate, 251. Starch, 575-577.
action of acids on (see under Conversion).
enzymes on
(see
under Conversion),
conversion
(see Conversion), determination by Fehling's solution, 438-442.
Sachsse's method, 439.

solution under pressure, 439. with diastase, 440, 441.
modification of Noyes, 442.

polariscopic methods, 506, 507. by solution under pressure, 506.
in
with hydrochloric acid, 506, 507. commercial dextrins, 509.
formation of isomaltose from, 706. maltose from, 683, 699, 706.
formula
of,
687.
hydrolysis (see under Conversion), microscopic appearance, 576.
molecular weight, 686, 687.

occurrence, 575, 576. preparation of, 576.
of d-glucose from, 580.
maltose from, 699.

properties, 576. soluble, Lintner's
method
for
making, 577.
Starch-conversion products, calculation of composition from specific rotation, 507. determination of moisture in, 26.
solution factors
Steffens's trisaccharate process, 678.
of, 31,
487.
relation to specific rotation, 487.
Stereopticon electric lamp, 152. Stolle's refractometer table, 62.
Ixxii
INDEX
Strohmer's refractometer table, 61, 62. Strontium bisaccharate, 679, 680.

process, 679, 680.
use in isolating sucrose, 647.
monosaccharate, 678, 679.

raffinosate, 739.
Strophanthin, 645. Strychnos alkaloids, use in resolving d, 1-acids, 786, 787. Sucrose, 645-682. absorption spectra with a-naphthol, 379.
resorcin, 381.
action of acids
upon (see Inversion). heat on solutions of, 656-659.
active
invertase upon (see under Invertase). and inactive molecules, 664.
boiling point of solutions, 651.

calorific value,
317-319.
compounds, 676-681.
concentration, influence on activity of invertase, 674.
specific rotation, 174-177.
saccharimeter readings, 118, 119.

configuration, 682. contraction of volume with water mixtures, 32-36. crystalline form, 647, 648.
influence of raffinose on, 735, 736.
decomposition by heat, 655-659. determination by chemical methods, 436-438.

direct polarization, 194r-262.
invert polarization, 263-281.

in presence of fructose
and
glucose, 485, 489.
raffinose,
282-286.
fermentations, 651-655. action of different yeasts, 714.
non-inverting organisms, 651.

freezing point of solutions, 325-330. high polarizing derivative, 658, 659.
influence
on action of invertase, 673, 674.

formation of osazones, 352, 353. reducing power of glucose, 427, 428.
inversion of (see Inversion), ions, hypothesis, of, 665.
melting point, 648. molecular weight determination, 322-332.
normal weight
(see
Normal
weight),
occurrence, 645-647. optically inactive derivative, 659.
osmotic pressure of solutions, 322-324.

plasmolysis
polarizing
by solutions of, 324, 325. power compared with quartz, 116, 117. preparation, from plant substances, 647.
TNDEX
Sucrose, preparation, manufacturing processes, 646. refining, 646, 647.
Ixxiii
preparation of d-fructose from, 617. d-glucose from, 581.

properties, 647, 648. protective action upon invertase, 675, 676. purification for standardizing saccharimeters, 121. reducing action upon Fehling's solution, 427, 428.
rotation dispersion
solubility, 648-650.
of, 116,
173.
in beet molasses, 649.
cane molasses, 650.

water, 649. influence of salts on, 649-650. specific gravity, 648. of solutions (see under Specific gravity),
specific rotation, 173-184, 651 (see also Specific rotation), technical processes for recovering, 678-680. temperature influence on saccharimetric determination, 126-130, 255-262. specific gravity of solutions, 30, 31.
specific rotation, 178, 179.
tests for, 681, 682.
value of Ventzke degree, 200, 201. verification of saccharimeters by means

viscosity of solutions, 307-313. Sucrose pipette, Spencer's, 205, 206.
of, 121, 122.
Sugar acids (see under Acids). Sugar alcohols (see under Alcohols). Sugar balance, 162. Sugar beets, colloidal water in, 229, 230.
determination of juice
in, in,
marc
227, 230. 228, 229.
sucrose
in,
227-246.
by
digestion with alcohol, 238, 239. Rapp-Degener method, 238. digestion with cold water, 239.
method, 240-242. method, 239. Sachs-Le Docte's method, 242-244. digestion with hot water, 240.
Kriiger's
Pellet's
Herzfeld's method, 244. Pellet's method, 240.
Sachs-Le Docte's method, 243, 244, expression of juice, 227-229. extraction with alcohol, 232-235.
Scheibler's
method, 233-235.
determination of sucrose in spent chips, 247.
by expression method, 247. Herzfeld's alcoholic digestion

sampling
of,
and extraction method, 247,
248.
225.
Ixxiv
INDEX
Sugar beets, sampling of, by Keil's boring rasp, 226. Sugar-beet molasses, composition of, 260, 649.
solubility of sucrose in, 649.
Sugar-beet products, composition
of,
260.
influence of temperature on polarization of, 258-260 Sugar cane, composition of mill pressings from 232. distribution of water in, 231.
determination of fibre
in,
248.
in,
sucrose
235-238.
by Zamaron's
determination of sucrose in bagasse
tissues of, 231.
of,
extractor, 235-238. 248, 249.

digestion, 248, 249.
by hot-water
Sugar cane molasses, composition
of, 259, 650. solubility of sucrose in, 650.
of,
Sugar cane products, composition Sugar flasks, 165-168. Sugar scale (see under Scales).
Sulphitation, 646.
259.
influence of temperature
on polarization
of,
258-261.
Sulphuric acid, color reactions of sugars with, 340, 341.

inverting power
of,
663.
Surface area of solution, influence on copper reduction, 419.
Sweet-water spindles, 47, 48. Sykes and Mitchell's method for determining diastatic power, 513.
Synanthrin, 615. Synthesis of sugars, by cyanhydrine reaction, 365, 366.
enzymic action, 704, 705, 718.

molecular rearrangement, 355, 625, 626. oxidation of alcohols, 770-772. reduction of lactones, 776, 777.
d-Tagatose, 626-628.
properties
and
tests, 627.
synthesis from d-galactose, 626, 627. 1-Tagatose, 628. d, 1-Tagatose, 628.
Takadiastase, 692, 693.

d-Talite, 611, 768.
tribenzal, 770.
d, 1-Talite, 768.
d-Talomucic
acid, 611.
d-Talonic acid, 611, 775.

d-Talose, 611, 612. action of different yeasts upon, 714. 1-Talose, 612.
Tannase, 573.
Tanret's researches on modifications of sugars, 191, 192.
d-galactose, 192, 603. d-glucose, 192, 581.
INDEX
Tanret's researches on modifications of lactose, 192, 710.
Ixxv
rhamnose, 192, 565.

Tartar emetic, 784. Tartaric acid, 784-787.
isomeric forms, 784. Pasteur's methods for resolving racemic acid, 784-787
Pasteur).
(see
under
Tartrate, sodium
ammonium,
785.
hemihedral forms of, 785. Technical methods of saccharimetry, 201-255. Temperature, adjustment of saccharimeters at variable, 129, 130.
coefficient for polarization of quartz, 126, 127. sucrose, 127.
sugars, 127-129.
saccharimeter readings, 255-262. specific rotation of sugars, 178-181.

corrections in Clerget's method, 264-269. determining fructose, 478.
galactose, 480. raffmose, 283, 284. refractive index, 64; Appendix, 21.
specific gravity, 30, 31
;
Appendix,
5, 16.
specific rotation, 178-181.
polarizing sugars, 255-262. errors in use of, 257-261.

for beet products, 260.
cane products, 259.
method
of
Browne, 258-261. U. S. Treasury Dept., 256, 257.
Wiley, 256. equations for specific rotation of sugars, 178-181.

influence on activity of diastase, 690, 691. invertase, 674.
pancreatin, 695, 696. copper-reducing power of sugars, 418.
length of polariscope tubes, 158. saccharimeter scales, 126. specific rotation of sugars, 178-181.
speed of inversion, 269, 664. viscosity of sugar solutions, 311. of optical inactivity of invert sugar, 287-289. polarization at constant (see under Polarization).
high (see under Polarization), regulation of refractometers, 58, 59, 73, 74.
water regulators for constant, 59, 60, 159, 160. Tetrasaccharides, 528, 574, 747-750. Tetroses, 540-543.
reaction for, 378.
Thiocyanate method
for determining copper, 414, 415. Thiosemicarb,".zone reaction of sugars, 366.
Ixxvi
d-Threose, 542.
1-Threose, 542. Time of boiling, influence
INDEX
upon copper reduction,
417, 418.
Tint polarimeters, 86, 88. Tint saccharimeters, 131, 132.

Tischtschenko's method for determining refractive index, 68, 69.
Tollens's "absatz"
method for studying absorption spectra, 344, 345. apparatus for hydrolysis, 548, 549.
vacuum evaporation, 549, 550. concentration formula for specific rotation of sucrose, 176. furfural reaction for pentose groups, 374, 375.
method
levulinic acid reaction for hexose groups, 372, 373. for determining galactose and galactan, 459, 460.
pentoses and pentosans, 449-453. naphthoresorcin test for pentoses and glucuronic acid, 383-385.
silver solution, 337.
Tollens and Ellett's
method for determining rhamnose and rhamnosan, 456, 457; Appendix, 89. Gans's saccharic acid test for glucose groups, 588. Krober's method for determining pentoses and pentosans, 449-452
;
Appendix, 83. Mayer's method for determining fucose and fucosan, 456, 457; Appendix,
89.
Oshima's test for methylfurfural, 386. Wheeler's method for preparing xylan, 553. Widtsoe's test for methylfurfural, 385. Yoder's test for dehydromucic acid, 781.
Tolman's modification of Clerget's method by inverting at ordinary temperature, 269.
Tolman and Smith's refractometer
table, 62, 63.
Total solids of sugar solutions, determination by methods of drying, 15-26. estimation from refractive index, 50-75.
specific gravity, 27-49.
results
compared by
different methods, 67-69.
Traganthose, 560. Trehabiose (see Trehalose).
Trehala-manna, 718, 719.

Trehalase, 720.
Trehalose, 718-721.
configuration, 721.
fermentation, .720.
occurrence, 718. preparation and properties, 719.
reactions, 719, 720. tests, 720, 721.
Trehalum, 718, 719. Trier for sampling sugar, 4-6. dimensions

Trihexose saccharides, 732-746.
Trimethyltrioses, 540. Trioses, 538, 539.
of, 5.
INDEX
Trioses, reactions for, 378.
Ixxvii
Trioxyglutaric acids, 529, 551, 556, 559.

Triple
field,
97, 98.
Trisaccharides, 528, 574, 731-746. Triticin, 615.
Tubes,
filter (see Filter
tubes), polariscope, 153-161. calibration of, 155, 156.
cover glasses and washers
for, 156.
desiccating caps for, 160, 161. expansion coefficients of, 158.
form of Landolt
(sliding caps), 157. Pellet (continuous polarization), 158, 159. Schmidt and Haensch (enlarged end), 157.
Yoder (volumetric),
mounting
of, 154,
161.
155.
of glass, 153-157.
metal, 157, 158.
with water-jacket, 158, 159. Tungstates, reaction with sugar alcohols, 766.
Tunicin, 579.
Turanose, 725, 726. formation from melezitose, 741.

hydrolysis, 725, 742.
preparation and properties, 725.

Unified copper-reduction methods, 424-426 (see under Copper-reduction). United States Coast Survey Sugar Scale, 114, 115.
United States Treasury Department regulations for sampling molasses,

temperature
256, 257.
10.
sugars, 5. corrections for saccharimeters,
Units of heat
(see Calories).
Units of volume
influence
(see
Cubic centimeter).
Urea, action upon sugars, 366.
on rate
of inversion, 272. rotation of fructose, glucose and invert sugar, 272.
use in Clerget's
method
of inversion, 271-273.
Ureide reaction of sugars, 366.
Vacuum, evaporation of sugar solutions in, 549, 550. methods of drying sugar products, 20-26. Van't Hoff and Le Bel's theory of the asymmetric carbon atom, Variability in reducing power of sugars, 400-402.
Vegetable-glycogen, 578. Velocity of inversion (see under Inversion). mutarotation (see under Mutarotation).
530, 758.
Ventzke sugar scale
(see
under Scales).
Verbascose, 749, 750. occurrence, 749.
Ixxviii
Verbascose, preparation, 749, 750.
INDEX
properties, 750. Verification of polarimeter readings (see under Polarimeters)
saccharimeter readings (see under Saccharimeters). Violette's volumetric method for determining reducing sugars, 393-395.
Viscosimeter, apparatus of Engler, 308.
principle of, 309 Viscosity, coefficient of, 309. of dextrin solutions, 508, 510.
sucrose solutions, 307-313.
diagram
of curves, 311. influence of concentration, 310. impurities, 311, 312.
temperature, 310, 311.

pipette, 307, 308. Viscous fermentation of sugars, 584, 652, 653. Volemite, 636, 768.
Volemose, 636, 637. Volquartz's hydrometer, 46, 47. Volume of precipitate error, 209-215. Volume, units of, 27, 28 (see Cubic centimeter). Volumetric methods for determining copper, 410-415.
reducing sugars, 389-399. conversion tables
for, 397, 398.
Volumetric polariscope tube, 161. Volumetric sugar flasks, 165-168. Vosatka's hydrometer, 47.
Walnut leaves, preparation of i-inosite from, Washers for polariscope tubes, 156.
Water, colloidal or imbibition, 229, 230, 246. digestion (see under Sugar beet), distribution in plant tissues, 230-232.
sugar cane, 231.
761.
Water-baths for constant temperature, 159, 160. Water-heater and pressure regulator, 59, 60.
Wave
length of light, 77.

of sugars, 173, 174. zero point of saccharimeters, 109, 110. length for sodium light, 116, 150. white light, 116.
influence
on rotation
Wave
theory of
Wedge system
light, 76, 77. of saccharimeters, 108-112.
control, 111, 112.
double, 110-112. single, 108-110.
working, 111, 112.
Weighing bottle
for solutions, 24. sugars, 16.
capsules, 16.
INDEX
Weighing dish for sugars, 203. Weight in vacuo, 38, 40, 164, 165. Weight, normal (see Normal weight). Wein's method for determining maltose, 423; Appendix, 40. Welsbach lamps, 152.
W'estphal's specific gravity balance, 40-42.
Ixxix
White
light,
lamps
for,
151-153.
lengths
of, 116.
mean wave
Wheeler and Tollens's method
for preparing xylan, 553. Wiesnegg's hot-air oven, 17, 18.
Wild's polaristrobometer, 98-100.
Wilhelmy's law of mass action, 660.

Wiley's desiccating caps for polariscope tubes, 160, 161. filter tube, 393.
method
for destroying optical activity of reducing sugars, 306.
determining dextrin, 306, 490.

fructose
298.
by
polarization at high temperature, 296-
temperature correction table for saccharimeters, 256. Wiley and EwelPs method for determining lactose in milk, 253.
Winter's cylinder for determining specific gravity, 45, 46.
Winton's lead number, 517, 518. Wohlgemuth's method for determining diastatic power, 515.
Wood gum (see Xylan). Wood sugar (see 1-Xylose).

Working wedge,
Worts, composition
111, 112. of, 690.
Xanthorhamnin, 564, 599.

Xylan, determination, 450-452; Appendix, 83
hydrolysis, 553, 554. occurrence, 553.
(see also
under Pentosans).
preparation, 553. properties, 553.

Xylite, 556, 767.
dibenzal, 770. oxidation to d, 1-xylose, 556. Xylogalactans, 599.
d, 1-Xyloketose, 562.
d-Xylonic acid, 552.
cadmium double
1-Xylonic acid, 555.
salt of, 552.
cadmium double
salt of, 555.
conversion to d-lyxonic acid, 775. oxidation to 1-threose, 542.

d-Xylose, 552. 1-Xylose, 552-556.
Bertrand's reaction
for, 555, 556.
Ixxx
1-Xylose, calorific value, 319.
INDEX
conversion to 1-gulose, 609.
1-idose, 611.
determination, 450-453; Appendix, 83
(see
under Pentoses).
in presence of 1-arabinose, 482.
d-glucose, 300, 301.
diformal, 556.
fermentation, 555. formation from d-glucuronic acid, 375, 532.

occurrence, 552554. oxidation to 1-xy Ionic acid, 556. with bromine, 363.
preparation from straw,
etc.,
554, 555.
xylan, 554.
properties, 555. reducing ratio to glucose, 421, 476. reduction to xylite, 556.
tests, 555, 556 (see also under Pentoses). value of Ventzke degree, 200, 201.
yield of furfural from, 449.
d,l-Xylose, 556. Xylo-proteids, 554.
Yeast, action of pure cultures on different sugars, 299, 714. autolysis of, 669.
extract, preparation of, 300. mannan, 594.
method
of O' Sullivan
and Tompson
for inversion, 274.
Ogilvie's modification, 274
microscopic appearance, 582.

non-inverting varieties
of,
651.
nutritive salt solution for, 299. occurrence of zymase in, 582.
preparation of invertase from, 669, 670.
maltoglucase from, 702. top- and bottom-fermentation varieties
of,
723.
action on melibiose, 723. raffinose, 738.
use in resolving racemic sugars, 787 Yoder's volumetric polariscope tube, 161.
(see also
under Saccharomyces)
Yoder and Tollens's
test for
dehydromucic
acid, 781.
Zamaron's hypochlorite method of
clarification, 218.
method
Zeiss's
of hot-water extraction, 235-238.

(see
immersion refractometer sodium lamp, 148, 149.
Immersion refractometer).
INDEX
Zeiss's spiral heater
Ixxxi
and water pressure regulator, 59, 60. Zero-point determination of polarimeters, 106, 107. saccharimeters, 109-112.
error for Bates's saccharimeter, 140-142. Zinc dust as a decolorizing agent, 278. Zinc nitrate method for decomposing saccharate, 251.
Zymase, 582. Zymogen, 683.
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A HANDBOOK OF

RESEARCH, TECHNICAL AND CONTROL LABORATORIES

Sugar Experiment Station, New Orleans,

NEW YORK JOHN WILEY & SONS

CHAPMAN & HALL,

Copyright, 1912, in Great Britain

GEH.-RATH PROF. DR.

so broad that in the selection of

Part II of the present volume, he has therefore included a brief

account of the occurrence, methods of preparation, properties and

sugars that the sugar analyst

N. Y., August, 1912.

The Simple Carbohydrates and

Guide pratique du Fabricant

Bujard and Baier

Beet Sugar Manufacture (1906).

Cane Sugar and its Manufacture Methods of Chemical Control

Gredinger Jago Konig....

Sugar Factories (1905). Die Raffination des Zuckers (1909).

of Bread Making (1911). Die Untersuchung landwirtschaftlich und

Technical Calculations for Sugar Works

Sykes and Ling.

Wiechmann Wiedemann and Ebert

Chemischen Methoden (1899). Principles and Practice of Agricultural

Bull. 107 of Chemistry.

Am. Chem. Jour Am. Sugar Ind

American Chemical Journal. American Sugar Industry.

Annalen der Chemie

Ann. chim. phys Archief Java Suiker Ind.. Archiv Pharm.

Biochem. Zeitschrift Bull, assoc. chim. sucr. dist.

Biochemische Zeitschrift. Bulletin de I'association des chimistes de sucrerie et de distillerie de France et

Centralblatt Centrbl. Zuckerind

Chemical News and Journal of Physical

prakt. Chem. J. Soc. Chem. Ind.

scient Zeitschrif t Oest.-Ung. Z. Zuckerind.

Monatshefte fiir Chemie. Moniteur scientifique.

Pogg. Ann Proceedings A. O. A.

Proceedings Int. Cong. App. Chem.

Sucrerie Beige West Indian Bull Wochenschr. f. Brauerei.

Chem Chem Chem

physiol. Chem Spiritusind

Unters. Nahr. Genussm.

des Vereins der Deutschen Zuckerindustrie. Zeitschrift fiir Zuckerindustrie in Bohmen.

PHYSICAL AND CHEMICAL METHODS OF SUGAR ANALYSIS

SAMPLING OF SUGAR AND SUGAR PRODUCTS

DETERMINATION OF MOISTURE IN SUGARS AND SUGAR PRODUCTS BY METHODS OF DRYING

V. POLARIZED LIGHT, VI.

THEORY AND DESCRIPTION OF POLARIMETERS

THEORY AND DESCRIPTION OF SACCHARIMETERS

METHODS OF SIMPLE POLARIZATION

X. METHODS OF INVERT OR DOUBLE POLARIZATION

METHODS OF SACCHARIMETRY 287 METHODS AS APPLIED TO THE EXAMINA307 333

XVII. MISCELLANEOUS APPLICATIONS

THE OCCURRENCE, METHODS OF PREPARATION, PROPERTIES AND PRINCIPAL

XIX. THE XX. THE XXI. THE XXII. THE

AMINO SUGARS AND THE CYCLOSES

XXIII. THE SUGAR ALCOHOLS AND SUGAR ACIDS

APPENDIX OF SUGAR TABLES

PHYSICAL AND CHEMICAL METHODS OF SUGAK ANALYSIS

SAMPLING OF SUGAR AND SUGAR PRODUCTS

best illustration of methods of sampling, is furnished by raw cane sugar.

of the errors con-